Team:XMU-China/Project

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Project

Bacterial chemotaxis, which is universal in E.coli, is defined as bacteria cells migration in response to a chemical stimulus. The natural E.coli chemotaxis has limited receptor proteins which can bind to only six kinds of amino acid. Nevertheless, the reprogrammed chemotaxis named pseudotaxis makes the bacteria able to respond to molecules, whose receptor proteins do not exist in classical E.coli, such as IPTG and L-arabinose, etc.

Figure 1 Chemotaxis mechanism of E.coli. The direction of rotation of the flagellar motor is controlled by the protein CheY. If the CheY is phosphorylated (CheY-P), it can bind to the flagellar motor protein FliM, causing the cell to tumble. While CheY is not phosphorylated, the flagellar motor rotates counterclockwise (CCW). [1]

E.coli have several flagella per cell (4–10 typically), which can rotate in two ways: counterclockwise (CCW) and clockwise (CW). [2] The former aligns the flagella into a single rotating bundle, causing the bacterium to swim in line, while the later breaks the flagella bundle apart such that each flagellum points in a different direction, causing the bacterium to tumble. The motility is determined by the phosphorylation state of CheY protein governed by CheZ protein. In the presence of CheZ protein, CheY-P is dephosphorylated and produce CheY, and the flagellar motor rotates CCW resulting in swimming. In the absence of CheZ, CheY is phosphorylated to CheY-P which binds to the flagellar switch protein FliM resulting in tumbling (Figure 1). [1] Therefore, we are able to control the bacterial motility by knocking out the CheZ gene of the wild-type then transfecting circuit containing CheZ gene into a CheZ knockout (ΔCheZ) strain. Besides, we introduce aptamers responding to a mass of specific molecules which can be applied to regulate gene expression, in our project, CheZ (Figure 2).

Figure 2 Mechanism of how aptamers controls the translation of CheZ protein. In the absence of target molecules (theophylline as an example). The mRNA’s ribosome binding site is paired, which inhibits the translation of CheZ protein. In the absence of CheZ, CheY-P will remain phosphorylated and the cells tumble in place. While in the presence of theophylline, the mRNA’s ribosome binding site will expose and the CheZ can be expressed, allowing the cells to run and tumble. [1]

Characterizing the circuit we constructed, we combine mathematical modeling with experiments, using modeling to guide experiments and to explain experimental phenomena. .

And we can characterize the efficiency of RBS and promoter via migration distance positively associated with the expression strength of CheZ.

, Mathematic is the simplest and clearest language, whose value to the development of human civilization is now widely recognized because its extensive application of science, society and daily life. However, the mathematical laws in life sciences is still unclear and even in chaos. Luckily, synthetic biology can overcome these shortcomings on some level. Based on this, we design a gene circuit and expect mathematical regularities to realize the regulation and control of life activities. We hope our work can inspire people's interests to combine mathematic with synthetic biology.

References

[1] Topp, Shana, and Justin P. Gallivan. "Guiding bacteria with small molecules and RNA." Journal of the American Chemical Society 129.21 (2007): 6807-6811.

[2] http://en.wikipedia.org/wiki/Chemotaxis

Overview

What means would you like to use to get a mathematical pattern? Draw one with compass and ruler, or type a function in a drawing software? Well, E.coli can help us to achieve our goals. We make the first attempt at introducing pseudotaxis of bacteria to form patterns in shape of conic section (such as ellipse and hyperbola). Firstly, let’s make precise mathematical definition on ellipse, hyperbola and parabola.

What are ellipse, hyperbola and parabola?

In mathematics, ellipse is a curve on a plane surrounding two focal points such that a straight line drawn from one of the focal points to any point on the curve and then back to the other focal point has the same length for every point on the curve (Figure 1).

Figure 1 Schematic of ellipse. Point F1, F2 are the two focal points and Point A is on the ellipse curve. The sum of the distance AF1 and AF2 is equal to the constant k: AF1+AF2=k.

And Hyperbola is a conic consisting of those points whose distances to some point, called a focus, and some line, called a directrix, are in a fixed ratio (>1), called the eccentricity. (Figure 2B)

Parabola is a conic whose eccentricity is equal to 1 (Figure 2C).

A

B

Figure 2A, Schematic of hyperbola. Point F1, F2 are the two focal points and Point A is on one of the hyperbola branches. The absolute value of the difference of the distance AF1, AF2 is equal to the constant k: |AF1-AF2|=k. 2B, one branch of hyperbola can be defined by directrix and eccentricity. Point A is a spot on the curve, point F is the focus. AB is the distance between A and directrix. The eccentricity e equal to TF/AB, and e>1.

Figure 2C Schematic of Parabola. Point A is a spot on the curve, point F is the focus. AB is the distance between A and directrix. The eccentricity e equals to AF/AB, and e=1.

Hypothesis

We find that the focal points, the constant and the ratio are the key of conic section. Therefore, we can easily get any eclipse by presetting reasonable focal points with the acceptable constant, as well as parabola or hyperbola by a fixed ratio. Based on these, we put forward the following hypothesis:

If we spot stimulus on semi-solid plate, it will spread from the spotting center out to the periphery. In the spreading process, the concentration is negative correlation to the distance from center. And the concentration gradient of the stimulus will be formed and maintained for a long period.
There is a threshold ratio of the concentrations of inducer and repressor. This means that more repressor will cause more repression, hence more inducer is needed to relieve the repression, and vice versa.

Design of circuit

Our circuit consists of two parts, of which one is named C (constraint), the other is named M (motile). (Figure 4):

We build our circuit in E.coli CL-1 which lacks gene lacI and CheZ (ΔlacI, ΔCheZ). At the absence of CheZ, CL-1 adopts non-motile phenotype.
Without any exogenous stimulus, E.coli will produce background amount of AraC to repress pBAD in limit degree. Even when L-arabinose isn’t involved in, promoter pBAD has expression leakage, so that part C will produces protein LacI which can bind to the operon of promoter pLac and thus repress its transcription. Because L-arabinose could induce pBAD, within certain concentration range, more L-arabinose involved in means that part C could produce more LacI to repress the expression of pLac. Because of its ability to constrain chemotaxis, this part is named C (abbreviate from constraint).
When IPTG is involved in, it can relieve the repression from LacI, therefore protein CheZ is produced to make our engineering bacteria (CL-1) regain motile ability. Within certain L-Arabinose concentration range which means certain constraint condition, more cheZ is produced with more IPTG involved in leading to stronger motile ability. Because of its ability to make CL-1 motile, this part is named M (abbreviate from motile).

Figure 4 Part C produces LacI to repress the expression of part M. Part M could produce CheZ to make CL-1 regain motile ability.

Characterization of circuit

We sequenced the circuit above and characterized it in E.coli CL-1. As CL-1 lacks LacI gene, promoter pLac won’t be repressed by background amount of LacI protein.

We apply gradient test to find out which influence would be made on reprogrammed chemotaxis under the following parameters: the concentration of chloramphenicol, IPTG and L-arabinose.

At first, we need to find out at which chloramphenicol concentration we could get the best chemotaxis performance. We try gradient concentration of chloramphenicol at semi-solid medium culture as Table 1 show. We find that the chemotaxis performance doesn’t have overt linear relationship to chloramphenicol. Interestingly, 50μg/ml of chloramphenicol gives CL-1 the best chemotaxis. So we applied that to our following characterization.

Characterization of circuit

We sequence the circuit above and characterize it in E.coli CL-1. As CL-1 lacks LacI gene, promoter pLac won’t be repressed by background amount of LacI protein.

We design gradient tests of the concentration of chloramphenicol, IPTG and L-arabinose to find out their influence on reprogrammed chemotaxis. To begin with, we test for the best chloramphenicol concentration. We test gradient concentration of chloramphenicol at semi-solid medium culture as Table 1 show. We find that the activity of chemotaxis doesn’t have overt linear relationship to chloramphenicol. Interestingly, 50μg/ml of chloramphenicol gives CL-1 the best chemotaxis. So we apply that to our following characterization.

Table 1 Curve of chemotaxis diameter under gradient concentration of Cm (chloramphenicol).

As promoter pBAD leads to a certain level of expression leakage of LacI, CL-1 has the worst chemotaxis. We added IPTG at gradient concentration and got the results (Table 2). We find that the chemotaxis performance keeps increasing when the concentration of IPTG increases from 0 and 0.02μM and gets the best performance with the IPTG range from 0.02 to 0.025μM. We apply 0.025μM IPTG for our following characterization.

Table 2 Curve of chemotaxis diameter over time under gradient concentration of IPTG.

As more L-arabinose added in, the expression from promoter pBAD will be stronger which leads to more LacI produced resulting in the inhibition to chemotaxis. As our expectation, chemotaxis performance keeps going down as the concentration of L-arabinose increases (Table 3). We find that 0.2% of L-arabinose has the best inhibitory effect on chemotaxis with 0.025μM of IPTG added in.

Table 3 Curve of chemotaxis diameter under gradient concentration of L-arabinose.

Eclipse

If we spot IPTG on the center of the semi-solid culture medium, concentration gradient will be formed as Figure 3A. Larger circle represents lower concentration with a lower number labeled.

A

B

Figure 3 Schematic of concentration gradient. A, single point spreading model. Larger number represents higher concentration while smaller number represents lower concentration. B, double points spreading model. Letter A~J represent equal concentration points on the ellipse with two labeled number added up to 6. Red curve represents the ellipse with two focal points on the IPTG spots.

If we spot two IPTG simultaneously, two spots’ concentration gradient will interact with each other to form the ellipse boundary as image in Figure 3B.

If we spot E.coli CL-1 with circuit above transformed in at the center between two IPTG spots on semi-solid culture plate as the image in Figure 5. The concentration of L-Arabinose added in the culture medium determines the degree of constraint from part C. Each red ellipse curve represents equal IPTG concentration, and there are a series of such ellipses from inside to outside represent IPTG concentrations from highest to lowest. One of the ellipses is the critical line indicating that the constraint from part C can just be relieved by the certain concentration of IPTG. Initially, as the concentration of IPTG is enough to relieve the constraint, bacteria can swim from the center out to periphery. When the bacteria swim out of the critical line, the concentration of IPTG can’t relieve the constraint, so the bacteria adopt non-motile phenotype. On the contrary, when the bacteria are inside of the critical line, they adopt motile phenotype. When the bacteria swim from inside to outside, motile bacteria become non-motile, so the bacteria will aggregate outside the critical line while the bacteria density inside the critical line will decline. Thus, an ellipse boundary is formed.

Figure 5 Schematic of Critical Line model for ellipse. Critical line represents the IPTG concentration which can just relieve the repression from part C. So that CL-1 is motile inside the ellipse while non-motile outside the ellipse.

Parabola and hyperbola

We got the optimum concentrations of IPTG and L-arabinose as inducer and repressor through preliminary experiments. Because the concentration of stimulus will decrease during spreading, so does their effect, we use IPTG and L-arabinose of which concentrations are a little bit higher than the optimum values for our experiments.

We draw a straight line with L-arabinose on the semi-solid culture medium, and a spot with the mixture of IPTG and CL-1 on one side of the line. In the area around the spot, the induction of IPTG is stronger than the repression of L-arabinose, CheZ is expressed and the bacteria adopt motile phenotype. However, when they approach the line where the repressor have a greater effect on the motility, they will lost their motile phenotype and stop.

According to the hypothesis 2, on the threshold ratio of the concentration of inducer and repressor, their effects are offset and a critical line is formed. Distances of the points on the critical line to the IPTG spot (focus), and the L-arabinose line (directrix) are in a fixed ratio (eccentricity). If the ratio is equal to 1, the critical line is a parabola. If the ratio is larger than 1, it is a branch of a hyperbola.

Figure 6 Schematic of Critical Line model for parabola and hyperbola. CL-1 becomes no-motile on the left side of the critical line while motile on the other side. The eccentricity e equals to PF/PB. If e=1, we define the critical line as parabola. If e>1, we define that as one branch of hyperbola.

Other function curves

After explorations, we found a new way to spot bacteria and stimulus and got a special function curves.

We draw two spots on the semi-solid culture medium, one with L-arabinose (Figure 7A) and the other with the mixture of IPTG and CL-1. Similarly, on the threshold ratio of the concentration of inducer and repressor, their effects are offset and a critical line is formed. Distances of the points on the critical line to the spot A and the spot B are in a fixed ratio (Figure 7B). Actually, as the critical line is quiet similar to hyperbola, we name it quasi-hyperbola.

A

B

Figure 7A Schematic of quasi-hyperbola formation on semi-solid medium culture. 7B Actual experiment result is shown.

Other interesting ways to spot bacteria and stimulus are waiting to be discovered, and the idea can be extended to other function curves and patterns.

Thousands of years ago in China, people began to preserve food by curing them which was recorded in Qimin Yaoshu around 540 AD (Figure 1). Curing is any of various food preservation and flavoring processes of foods such as meat, fish and vegetables, by the addition of a combination of salt, nitrates, nitrite or sugar and it is one of the oldest methods of preserving food. Table salt is the primary ingredient used in food curing. Removal of water and addition of salt to meat creates a solute-rich environment where osmotic pressure draws water out of microorganisms, slowing down their growth. Doing this requires a concentration of salt of nearly 20%. It has already been proved that 5% concentration of NaCl could inhibit the growth of E.coli. However, utilizing hyperosmotic pressure to kill E.coli

haven’t been fully explored in synthetic biology. This year, our team have put efforts on this topic and developed a system that will contribute to biosafety.

Figure 1 The production process of curing food.

Figure 2 The schematic of osmotic-taxis design.

E.coli makes use of the EnvZ/OmpR system to mediate signal transduction in response to environmental osmolarity changes. EnvZ, a histidine kinase, undergoes trans-autophosphorylation, then the high-energy phosphoryl group is subsequently transferred to OmpR, a response regulator.

In our system, we involved OmpR-controlled promoter (PompC) in (Figure 2). The expression strength of PompC is depending upon the medium osmolarity. When medium osmolarity is increasing, the EnvZ will phosphorylate more OmpR into phosphorylated OmpR (OmpR-P), resulting in stronger expression strength of PompR. In our circuitry design, CheZ is upstream regulated by PompR.

We use semi-solid medium culture with gradient concentration of sucrose to characterize the device (BBa_K1412008). And we assume that the motile ability is proportional to the moving radius. In the plot (Figure 3), when no sucrose added in, the motile ability is the weakest. The motile ability keeps growing as the concentration of sucrose increases from 0 to 4%. Then the motile ability goes down slightly as the sucrose concentration increased from 4% to 10%, but the ability is still stronger than that at concentration 0. We can draw a conclusion that our device is working as expectation, the motile ability goes down (4%~10%) because of the inhibition from hyperosmotic pressure.

Figure 3 The plot of moving radius versus sucrose concentration. The four curves were measured after 10h, 11h, 12h and 16.5h respectively.

Based on the characterization, we spotted hyperosmotic pressure spot and reprogrammed CL-1 spot on semi-solid medium culture as Figure 4 shows. The concentration will decrease with the increase of the distance away from hyperosmotic pressure spot. As the osmotic pressure is proportional to the medium concentration. The moving tendency of reprogrammed CL-1 will orient to the hyperosmotic pressure spot. Even at the inhibiting osmotic pressure, the motile ability is still stronger than that without any inducer. So reprogrammed CL-1 may even swim towards the high-osmotic site and die. The killing mechanism is just like the black hole. When the bacteria move into the “event horizon” where the osmotic pressure reaches to the critical value named the killing osmotic pressure, the bacteria can’t go out of the border

and be killed finally.

Figure 4 Schematic of killing bacteria by black hole.

It’s very cheap and accessible to get the source (such as NaCl and sucrose) to create hyperosmotic pressure, while antibiotics is expensive and have a bad effect on environmental microbiology because of drug resistance. However, the source to hyperosmotic pressure is environmentally friendly and won’t generate the risk of drug resistance. If our black hole system could be fully developed, it will reduce the access barriers to microbiology research especially for the scientists from poor countries.

The sources (such as NaCl and sucrose) to create hyperosmotic pressure are cheap, accessible and environmentally friendly, while antibiotics is expensive and have a bad effect on environmental microbiology because of drug resistance. If our black hole system could be fully developed, barriers to microbiology research will be removed especially for the scientists from poor countries.

XMU-China 2013 has tried to construct oscillation system by standard biobricks. The synchronized oscillation system used in that study (Figure 1A) is based on the quorum sensing machineries in Vibrio fischeri and Bacillus thurigensis. Three identical luxI promoters are in charge of luxI (from V. fischeri), aiiA (from B.thurigensis) and gfp genes separately. The LuxI synthase generates an acyl-homoserine-lactone (AHL), which can spread across the cell membrane and mediate intercellular coupling. AHL then binds to LuxR produced intracellularly, and the LuxR-AHL complex would activate the luxI promoter. AiiA catalyzes the degradation of AHL as the negative feedback in the circuit. Therefore, both the activator AHL and the repressor AiiA of the network are activated by the luxI promoter simultaneously.

A

B

Figure 1 A

B Two oscillation cycles were observed within 500 minutes.

Based on above principle, one published paper has already realized synchronized oscillations under microfluidic device. However, XMU-China 2013 can’t get synchronized oscillation on microfluidics, and that will be discussed later. Through calculating fluorescence on 96-microwell plate every 15 minutes, they got two oscillation cycles within 500 minutes (Figure 1B).

Based on that, we construct our circuit by replacing GFP with CheZ (Figure 2). As the expression strength of CheZ is oscillatory fluctuating, the motile ability will change periodically. Bacteria will have the strongest motile ability at wave crest while even be non-motile at wave trough. Thus, the periodical change of motile ability leads to bacteria density distributing unevenly. When the bacteria are at non-motile period, they will aggregate together leading to the formation of growth-ring-like patterns which could be distinguished by naked eyes.

Many trees in temperate zones make one growth ring each year, with the newest adjacent to the bark. We can tell a tree’s age by counting the number of growth rings. Analogously, bacteria rings could also be formed by gene oscillator. Multiply the period by the quantity of bacteria rings, we can tell how much time has passed.

Figure 2

In the project of iGEM13 XMU-China, they can’t get expected oscillation. However, this year iGEM14 XMU-China further investigate the reason of abnormal oscillation. We further review SDS-PAGE analysis to confirm the circuit at protein level. The SDS-PAGE data is shown in Figure 1. Based on that, we make a reasonable assumption that the unexpected behavior of the LuxR Promoter leads to the misfolding proteins hence the abnormal oscillation.

Figure 1 SDS-PAGE analysis of E.coli K strain (DH5α). (a) Lane 1-2: supernatant and pellet of original DH5α; Lane 3-4: supernatant and pellet of strain with single plasmid A1 (BBa_K1036003); Lane 5-6: supernatant and pellet of strain with both plasmids A1 (BBa_K1036003) and B (BBa_K1036000). The red arrows indicate the misfolding GFP-LVA protein (27.6 kDa) in the precipitation. (b) Lane 1-2: supernatant and pellet of original BL21; Lane 3-4: supernatant and pellet of strain with single plasmid A1 (BBa_K1036003); Lane 5-6: supernatant and pellet of strain with both plasmids A1 (BBa_K1036003) and B (BBa_K1036000). The blue arrows indicate LuxR (27.5 kDa), GFP-LVA (27.6 kDa) and AiiA-LVA (28.7 kDa) in the supernatant. The orange arrows indicate LuxI-LVA (22.4 kDa) in the supernatant. (The marker of b was not in right position, however, the proteins were confirmed by MALDI-TOF-TOF .)

As the SDS-PAGE shows, a large amount of GFP-LVA and LuxI-LVA appear in pellet where misfolding proteins often exist. Both proteins directly affect the oscillation result. And it is critical to find out the reason for misfolding proteins. iGEM14 XMU-China make the following assumption:

The 2012 published paper reveals an unexpected behavior of Lux pR (BBa_R0062). In the absence of autoinducer 3OC6 (AHL), LuxR binds to Plux (Lux pR) and activates backwards transcription (Figure 2).

Figure 2 Relative RFP fluorescence for a control construct designed to measure backwards transcription from Lux pR. Addition of LuxR and 3OC690 (AHL) as indicated. Error bars in all panels are one standard deviation.

The imperfect simplification of setting lux pL and Lux pR in the same direction:

From the original design by Jeff Hasty, we can see that Lux pR and Lux pL are set in opposite directions (Figure 3A). In the absence of AHL, LuxR could activate backward transcription of Lux pR leading to more expression of LuxR which is critical to meet the oscillation conditions. However, present literature don’t consider the backwards transcription which have effect on quorum sensing oscillation.

A. Original Design

B. iGEM13 XMU-China Design

Figure 3 A. Top row is the original design by Jeff Hasty. B. Bottom row is the simplified design which sets lux pL and lux pR in the same direction.

In the simplified design (Figure 3B), when LuxR activates the backward transcription, RNA polymerase will be blocked by the terminators B0015. So that this simplification doesn’t follow the original design. Actually, the reverse terminated efficiency of B0015 is 0.295(CC) which may lead to leakage transcription. However, the correct sequence of GFP-LAA can’t be transcribed during the backwards transcription, even if the minus-strand of GFP-LAA could be transcribed, the sequence of the RNA is not the right codons of GFP-LAA. hence incorrect amino acid sequences may be translated, resulting in misfolding GFP expression which is just as the SDS-PAGE shows (Figure 1). Misfolding GFP may also be produced by bi-directional transcription.

Because of the imperfect simplified design doesn’t follow the original function completely, the abnormal oscillation is justifiable. The misfolding protein is a evidence to support our assumption.

iGEM14 XMU-China involved sequence comparison to investigate the difference between the original and the registry parts. We find that the original Lux pR has 20bp overlapping sequence with original Lux pR. There is a restriction enzyme cutting site (EcoR I) at the 56bp of original Lux pR (Figure 4).

Figure 4 Schematic of original P system.

Parts registry truncate the original Lux pR at 56bp to get the 55bp Lux pR (BBa_R0062). On the contrary, Lux pL (BBa_R0063) is longer the original Lux pL, and at the end of BBa_R0063 is initial part of 41bp LuxR (BBa_C0062). Thus new problems arise—Is the modification of original P system reasonable? Does the modification result in the unexpected backward transcription?

Quorum sensing system is so widely used in the synthetic biology, we think it’s remarkable to make it clear. We highlight the abnormal phenomenon of QS oscillation which may be caused by imperfect simplification for the very first time. We hope that more effort would be made to figure out the interaction between QS oscillation parts.

Team:XMU-China/Project

From 2014.igem.org

Latest revision as of 18:32, 14 October 2014

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+			.left-menu li a {
+			    padding: 0px 10px 0px 0px;
+			}
+			.left-menu .selected {
+			#    color: white;
+			}
+			#.left-menu .selected:hover {
+			#    color: #5555FF;
+			#}
+			.left-menu:hover a {
+			    color: white;
+			}
+			.right-menu li {
+			#    background-color: #606060;
+			}
+			.right-menu li a {
+			    padding: 0px 15px 0px 0px;
+			    color: white;
+			    background-color: #606060;
+			}
+			.right-menu li a:hover {
+			    color: #aaaaff;
+			    text-decoration: underline;
+			}
 		</style>
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-								<li><a href="#">Background</a></li>
+								<li><a href="#Project_background">Background</a></li>
-								<li><a href="#">Design</a></li>
+								<li><a href="#Project_Circuits_Design">Design</a></li>
-								<li><a href="#">Modelling</a></li>
+								<li><a href="#Project_">Modelling</a></li>
 								<li><a href="#">Results</a></li>
 								<li><a href="#">Application</a></li>
@@ Line 359: / Line 437: @@
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 						<li id="navbar_main_02" class="navbar_main">
-							<a href="#"><span class="nav_bar" id="nav_bar_02"> </span></a>
+							<a href="https://2014.igem.org/Team:XMU-China/p&p"><span class="nav_bar" id="nav_bar_02"> </span></a>
-							<a href="#" class="nav_bar_full" id="nav_bar_full_02">Notebook</a>
+							<a href="https://2014.igem.org/Team:XMU-China/p&p" class="nav_bar_full" id="nav_bar_full_02">P&amp;P</a>
 							<ul class="navbar_sub">
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@@ Line 370: / Line 448: @@
 						</li>
 						<li id="navbar_main_03" class="navbar_main">
-							<a href="#"><span class="nav_bar" id="nav_bar_03"> </span></a>
+							<a href="https://2014.igem.org/Team:XMU-China/judging"><span class="nav_bar" id="nav_bar_03"> </span></a>
-							<a href="#" class="nav_bar_full" id="nav_bar_full_03">Judging</a>
+							<a href="https://2014.igem.org/Team:XMU-China/judging" class="nav_bar_full" id="nav_bar_full_03">Judging</a>
 						</li>
 						<li id="navbar_main_04" class="navbar_main">
-							<a href="#"><span class="nav_bar" id="nav_bar_04"> </span></a>
+							<a href="https://2014.igem.org/Team:XMU-China/notebook"><span class="nav_bar" id="nav_bar_04"> </span></a>
-							<a href="#" class="nav_bar_full" id="nav_bar_full_04">Team</a>
+							<a href="https://2014.igem.org/Team:XMU-China/notebook" class="nav_bar_full" id="nav_bar_full_04">Notebook</a>
 							<ul class="navbar_sub">
 								<img class="navbar_sub_x" id="navbar_sub_x_04" src="https://static.igem.org/mediawiki/2014/1/1f/Xmu_navbar_sub_04.png" />
-								<li><a href="#">Lab notes</a></li>
+								<li><a href="https://2014.igem.org/Team:XMU-China/notebook#labNote">Lab notes</a></li>
-								<li><a href="#">Calendar</a></li>
+								<li><a href="https://2014.igem.org/Team:XMU-China/notebook#caption">Calendar</a></li>
 							</ul>
 						</li>
 						<li id="navbar_main_05" class="navbar_main">
-							<a href="#"><span class="nav_bar" id="nav_bar_05"> </span></a>
+							<a href="https://2014.igem.org/Team:XMU-China/team"><span class="nav_bar" id="nav_bar_05"> </span></a>
-							<a href="#" class="nav_bar_full" id="nav_bar_full_05">P&amp;P</a>
+							<a href="https://2014.igem.org/Team:XMU-China/team" class="nav_bar_full" id="nav_bar_full_05">Team</a>
 							<ul class="navbar_sub">
 								<img class="navbar_sub_x" id="navbar_sub_x_05" src="https://static.igem.org/mediawiki/2014/d/de/Xmu_navbar_sub_05.png" />
-								<li><a href="#">Members</a></li>
+								<li><a href="https://2014.igem.org/Team:XMU-China/team#teamMember">Members</a></li>
-								<li><a href="#">Gallery</a></li>
+								<li><a href="https://2014.igem.org/Team:XMU-China/team#ourTeam">Gallery</a></li>
 							</ul>
 						</li>
 						<li id="navbar_main_06" class="navbar_main">
-							<a href="#"><span class="nav_bar" id="nav_bar_06"> </span></a>
+							<a href="https://2014.igem.org/Team:XMU-China/safety"><span class="nav_bar" id="nav_bar_06"> </span></a>
-							<a href="#" class="nav_bar_full" id="nav_bar_full_06">Safety</a>
+							<a href="https://2014.igem.org/Team:XMU-China/safety" class="nav_bar_full" id="nav_bar_full_06">Safety</a>
 						</li>
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@@ Line 413: / Line 491: @@
 					</li>
 					<li class="sidebar_main">
-						<h3><a href="#">P&P</a></h3>
+						<h3><a href="https://2014.igem.org/Team:XMU-China/p&p">P&P</a></h3>
 						<ul class="sidebar_sub">
 							<li><a href="#">Human Practice</a></li>
@@ Line 422: / Line 500: @@
 					</li>
 					<li class="sidebar_main" id="side_bar_judging">
-						<h3><a href="#">Judging</a></h3>
+						<h3><a href="https://2014.igem.org/Team:XMU-China/judging">Judging</a></h3>
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@@ Line 428: / Line 506: @@
 				<ul id="side_bar_main_2">
 					<li class="sidebar_main">
-						<h3><a href="#">Notebook</a></h3>
+						<h3><a href="https://2014.igem.org/Team:XMU-China/notebook">Notebook</a></h3>
 						<ul class="sidebar_sub">
 							<li><a href="#">Lab notes</a></li>
@@ Line 435: / Line 513: @@
 					</li>
 					<li class="sidebar_main">
-						<h3><a href="#">Team</a></h3>
+						<h3><a href="https://2014.igem.org/Team:XMU-China/team">Team</a></h3>
 						<ul class="sidebar_sub">
 							<li><a href="#">Members</a></li>
@@ Line 442: / Line 520: @@
 					</li>
 					<li class="sidebar_main" id="side_bar_safety">
-						<h3><a href="#">Safety</a></h3>
+						<h3><a href="https://2014.igem.org/Team:XMU-China/safety">Safety</a></h3>
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 				</ul>
@@ Line 451: / Line 529: @@
 				<div id="Project_background_wd">
 					<p style="line-height: 150%;">
-					    <span style="line-height: 150%; font-size: 27px;"><span style="font-family: Times New Roman;">&nbsp;</span></span>
+					    <span style="line-height: 150%; font-size: 27px;"><span>&nbsp;</span></span>
 					</p>
 					<p style="text-align: justify;">
-					    <span style="font-family: Times New Roman;">Bacterial chemotaxis, which is universal in <em>E.coli</em>, is defined as bacteria cells migration in response to a chemical stimulus. The natural <em>E.coli </em>chemotaxis has limited receptor proteins which can bind to only six kinds of amino acid. Nevertheless, the reprogrammed chemotaxis named pseudotaxis makes the bacteria able to respond to molecules, whose receptor proteins do not exist in classical <em>E.coli</em>, such as IPTG and L-arabinose, etc. </span>
+					    <span>Bacterial chemotaxis, which is universal in <em>E.coli</em>, is defined as bacteria cells migration in response to a chemical stimulus. The natural <em>E.coli </em>chemotaxis has limited receptor proteins which can bind to only six kinds of amino acid. Nevertheless, the reprogrammed chemotaxis named pseudotaxis makes the bacteria able to respond to molecules, whose receptor proteins do not exist in classical <em>E.coli</em>, such as IPTG and L-arabinose, etc. </span>
 					</p>
 					<p>
-					    <span style="font-family: Times New Roman;">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
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@@ Line 464: / Line 542: @@
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 					                <p>
-					                    <span style="font-family: Times New Roman;"> &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;
+					                    <span> &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;
 					                    	<img width="600px" style="" src="https://static.igem.org/mediawiki/2014/a/a7/Xmu_project_background01.png"/>
 					                    </span>
@@ Line 473: / Line 551: @@
 					            <td width="553" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p>
-					                    <span style="font-size: 12px;"><strong><span style="font-family:;" new="" times="">Figure </span></strong><strong><span style="font-family:;" new="" times="">1</span></strong><strong> </strong><span style="font-family:;" new="" times="">Chemotaxis &nbsp; mechanism of<em> E.coli. </em>The direction &nbsp; of rotation of the flagellar motor is controlled by the protein CheY. If the &nbsp; CheY is phosphorylated (CheY-P), it can bind to the flagellar motor protein &nbsp; FliM, causing the cell to tumble. While CheY is not phosphorylated, the &nbsp; flagellar motor rotates counterclockwise (CCW). [1]</span></span>
+					                    <span style="font-size: 12px;"><strong><span>Figure </span></strong><strong><span>1</span></strong><strong> </strong><span>Chemotaxis &nbsp; mechanism of<em> E.coli. </em>The direction &nbsp; of rotation of the flagellar motor is controlled by the protein CheY. If the &nbsp; CheY is phosphorylated (CheY-P), it can bind to the flagellar motor protein &nbsp; FliM, causing the cell to tumble. While CheY is not phosphorylated, the &nbsp; flagellar motor rotates counterclockwise (CCW). [1]</span></span>
 					                </p>
 					            </td>
@@ Line 480: / Line 558: @@
 					</table>
 					<p>
-					    <span style="color: red;"><span style="font-family: Times New Roman;">&nbsp;</span></span>
+					    <span style="color: red;"><span>&nbsp;</span></span>
 					</p>
 					<p style="text-align: justify;">
-					    <span style="font-family: Times New Roman;"><em>E.coli</em> have several flagella per cell (4–10 typically), which can rotate in two ways: counterclockwise (CCW) and clockwise (CW). [2] The former aligns the flagella into a single rotating bundle, causing the bacterium to swim in line, while the later breaks the flagella bundle apart such that each flagellum points in a different direction, causing the bacterium to tumble. The motility is determined by the phosphorylation state of CheY protein governed by CheZ protein. In the presence of CheZ protein, CheY-P is dephosphorylated and produce CheY, and the flagellar motor rotates CCW resulting in swimming. In the absence of CheZ, CheY is phosphorylated to CheY-P which binds to the flagellar switch protein FliM resulting in tumbling (Figure 1). [1] Therefore, we are able to control the bacterial motility by knocking out the <em>CheZ</em> gene of the wild-type then transfecting circuit containing <em>CheZ</em> gene into a <em>CheZ</em> knockout (Δ<em>CheZ</em>) strain. Besides, we introduce aptamers responding to a mass of specific molecules which can be applied to regulate gene expression, in our project,<em> CheZ </em><strong>(Figure 2)</strong>. </span>
+					    <span><em>E.coli</em> have several flagella per cell (4–10 typically), which can rotate in two ways: counterclockwise (CCW) and clockwise (CW). [2] The former aligns the flagella into a single rotating bundle, causing the bacterium to swim in line, while the later breaks the flagella bundle apart such that each flagellum points in a different direction, causing the bacterium to tumble. The motility is determined by the phosphorylation state of CheY protein governed by CheZ protein. In the presence of CheZ protein, CheY-P is dephosphorylated and produce CheY, and the flagellar motor rotates CCW resulting in swimming. In the absence of CheZ, CheY is phosphorylated to CheY-P which binds to the flagellar switch protein FliM resulting in tumbling (Figure 1). [1] Therefore, we are able to control the bacterial motility by knocking out the <em>CheZ</em> gene of the wild-type then transfecting circuit containing <em>CheZ</em> gene into a <em>CheZ</em> knockout (Δ<em>CheZ</em>) strain. Besides, we introduce aptamers responding to a mass of specific molecules which can be applied to regulate gene expression, in our project,<em> CheZ </em><strong>(Figure 2)</strong>. </span>
 					</p>
 					<p>
-					    <span style="font-size: 19px;"><span style="font-family: Times New Roman;">&nbsp;</span></span>
+					    <span style="font-size: 19px;"><span>&nbsp;</span></span>
 					</p>
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@@ Line 493: / Line 571: @@
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 					                <p>
-					                    <img width="600"style="position:relative;margin-left:-300px;left:50%" src="https://static.igem.org/mediawiki/2014/2/27/Xmu_project_background02.png" />
+					                    <img width="600"style="position:relative;" src="https://static.igem.org/mediawiki/2014/2/27/Xmu_project_background02.png" />
 					                </p>
 					            </td>
@@ Line 500: / Line 578: @@
 					            <td width="553" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p>
-					                    <span style="font-size: 12px;"><strong><span style="font-family:;" new="" times="">Figure </span></strong><strong><span style="font-family:;" new="" times="">2</span></strong><strong> </strong><span style="font-family:;" new="" times="">&nbsp;Mechanism of how aptamers controls the &nbsp; translation of CheZ protein. In the absence of target molecules (theophylline &nbsp; as an example). The mRNA’s ribosome binding site is paired, which inhibits &nbsp; the translation of CheZ protein. In the absence of CheZ, CheY-P will remain &nbsp; phosphorylated and the cells tumble in place. While in the presence of &nbsp; theophylline, the mRNA’s ribosome binding site will expose and the CheZ can &nbsp; be expressed, allowing the cells to run and tumble. [1]</span></span>
+					                    <span style="font-size: 12px;"><strong><span>Figure </span></strong><strong><span>2</span></strong><strong> </strong><span>&nbsp;Mechanism of how aptamers controls the &nbsp; translation of CheZ protein. In the absence of target molecules (theophylline &nbsp; as an example). The mRNA’s ribosome binding site is paired, which inhibits &nbsp; the translation of CheZ protein. In the absence of CheZ, CheY-P will remain &nbsp; phosphorylated and the cells tumble in place. While in the presence of &nbsp; theophylline, the mRNA’s ribosome binding site will expose and the CheZ can &nbsp; be expressed, allowing the cells to run and tumble. [1]</span></span>
 					                </p>
 					            </td>
@@ Line 507: / Line 585: @@
 					</table>
 					<p>
-					    <span style="font-size: 19px;"><span style="font-family: Times New Roman;">&nbsp;</span></span>
+					    <span style="font-size: 19px;"><span>&nbsp;</span></span>
 					</p>
 					<p style="text-align: justify;">
-					    <span style="font-family: Times New Roman;">Characterizing the circuit we constructed, we combine mathematical modeling with experiments, using modeling to guide experiments and to explain experimental phenomena. .</span>
+					    <span>Characterizing the circuit we constructed, we combine mathematical modeling with experiments, using modeling to guide experiments and to explain experimental phenomena. .</span>
 					</p>
 					<p style="text-align: justify;">
-					    <span style="font-family: Times New Roman;">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p style="text-align: justify;">
-					    <span style="font-family: Times New Roman;">And we can characterize the efficiency of RBS and promoter via migration distance positively associated with the expression strength of <em>CheZ</em>.</span>
+					    <span>And we can characterize the efficiency of RBS and promoter via migration distance positively associated with the expression strength of <em>CheZ</em>.</span>
 					</p>
 					<p>
-					    <span style="font-size: 19px;"><span style="font-family: Times New Roman;">&nbsp;</span></span>
+					    <span style="font-size: 19px;"><span>&nbsp;</span></span>
 					</p>
 					<p style="text-align: justify;">
-					    <span style="font-family: Times New Roman;">, Mathematic is the simplest and clearest language, whose value to the development of human civilization is now widely recognized because its extensive application of science, society and daily life. However, the mathematical laws in life sciences is still unclear and even in chaos. Luckily, synthetic biology can overcome these shortcomings on some level. Based on this, we design a gene circuit and expect mathematical regularities to realize the regulation and control of life activities. We hope our work can inspire people&#39;s interests to combine mathematic with synthetic biology.</span>
+					    <span>, Mathematic is the simplest and clearest language, whose value to the development of human civilization is now widely recognized because its extensive application of science, society and daily life. However, the mathematical laws in life sciences is still unclear and even in chaos. Luckily, synthetic biology can overcome these shortcomings on some level. Based on this, we design a gene circuit and expect mathematical regularities to realize the regulation and control of life activities. We hope our work can inspire people&#39;s interests to combine mathematic with synthetic biology.</span>
 					</p>
 					<p>
-					    <span style="font-size: 19px;"><span style="font-family: Times New Roman;">&nbsp;</span></span>
+					    <span style="font-size: 19px;"><span>&nbsp;</span></span>
 					</p>
 					<p>
-					    <strong><span style="font-family: 宋体; font-size: 19px;">参考文献</span></strong>
+					    <strong><span style="font-family: 宋体; font-size: 19px;">References</span></strong>
 					</p>
 					<p style="text-align: justify;">
-					    <span style="font-family: Times New Roman;">[1] Topp, Shana, and Justin P. Gallivan. &quot;Guiding bacteria with small molecules and RNA.&quot; Journal of the American Chemical Society 129.21 (2007): 6807-6811.</span>
+					    <span>[1] Topp, Shana, and Justin P. Gallivan. &quot;Guiding bacteria with small molecules and RNA.&quot; Journal of the American Chemical Society 129.21 (2007): 6807-6811.</span>
 					</p>
 					<p style="text-align: justify;">
-					    <span style="font-family: Times New Roman;">[2] http://en.wikipedia.org/wiki/Chemotaxis</span>
+					    <span>[2] http://en.wikipedia.org/wiki/Chemotaxis</span>
 					</p>
 					<p>
-					    <span style="font-size: 19px;"><span style="font-family: Times New Roman;">&nbsp;</span></span>
+					    <span style="font-size: 19px;"><span>&nbsp;</span></span>
 					</p>
 					<p>
@@ Line 552: / Line 630: @@
 					</p>
 					<p>
-					    <span style="color: rgb(0, 112, 192); font-family:;" new="" times="">What means would you like to use to get a mathematical pattern? Draw one with compass and ruler, or type a function in a drawing software? Well, <em>E.coli</em> can help us to achieve our goals. We make the first attempt at introducing pseudotaxis of bacteria to form patterns in shape of conic section (such as ellipse and hyperbola).</span><span style="color: rgb(0, 112, 192);"> </span><span style="font-family:;" new="" times="">Firstly, let’s make precise mathematical definition on ellipse, hyperbola and parabola.</span>
+					    <span style="color: rgb(0, 112, 192); font-family:;" new="" times="">What means would you like to use to get a mathematical pattern? Draw one with compass and ruler, or type a function in a drawing software? Well, <em>E.coli</em> can help us to achieve our goals. We make the first attempt at introducing pseudotaxis of bacteria to form patterns in shape of conic section (such as ellipse and hyperbola).</span><span style="color: rgb(0, 112, 192);"> </span><span>Firstly, let’s make precise mathematical definition on ellipse, hyperbola and parabola.</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <strong><span style="font-family:;" new="" times="">What are ellipse, hyperbola and parabola?</span></strong>
+					    <strong><span>What are ellipse, hyperbola and parabola?</span></strong>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">In mathematics, ellipse is a curve on a plane surrounding two focal points such that a straight line drawn from one of the focal points to any point on the curve and then back to the other focal point has the same length for every point on the curve (<strong>Figure 1</strong>).</span>
+					    <span>In mathematics, ellipse is a curve on a plane surrounding two focal points such that a straight line drawn from one of the focal points to any point on the curve and then back to the other focal point has the same length for every point on the curve (<strong>Figure 1</strong>).</span>
 					</p>
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 					                <p>
-					                    <span style="font-family:;" new="" times=""> &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;
+					                    <span> &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;
 					                    	<img width="600" src="https://static.igem.org/mediawiki/2014/e/eb/Xmu_project_CircuitsDesign01.png"/>
 					                    </span>
@@ Line 580: / Line 658: @@
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 					                <p>
-					                    <strong><span style="font-family:;" new="" times="">Figure 1 </span></strong><span style="font-family:;" new="" times="">Schematic of &nbsp; ellipse. Point F1, F2 are the two focal points and Point A is on the ellipse &nbsp; curve. The sum of the distance AF1 and AF2 is equal to the constant k: &nbsp; AF1+AF2=k.</span>
+					                    <strong><span>Figure 1 </span></strong><span>Schematic of &nbsp; ellipse. Point F1, F2 are the two focal points and Point A is on the ellipse &nbsp; curve. The sum of the distance AF1 and AF2 is equal to the constant k: &nbsp; AF1+AF2=k.</span>
 					                </p>
 					            </td>
@@ Line 587: / Line 665: @@
 					</table>
 					<p>
-					    <span style="font-family:;" new="" times="">And Hyperbola is a conic consisting of those points whose distances to some point, called a focus, and some line, called a directrix, are in a fixed ratio (&gt;1), called the eccentricity. (<strong>Figure 2B</strong>)</span>
+					    <span>And Hyperbola is a conic consisting of those points whose distances to some point, called a focus, and some line, called a directrix, are in a fixed ratio (&gt;1), called the eccentricity. (<strong>Figure 2B</strong>)</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">Parabola is a conic whose eccentricity is equal to 1 (<strong>Figure 2C</strong>).</span>
+					    <span>Parabola is a conic whose eccentricity is equal to 1 (<strong>Figure 2C</strong>).</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span><span style="font-family:;" new="" times="">&nbsp;</span><span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span><span>&nbsp;</span><span>&nbsp;</span>
 					</p>
 					<table cellspacing="0" cellpadding="0" width="600px" style="position:relative;margin-left:-300px;left:50%">
@@ Line 600: / Line 678: @@
 					            <td width="302" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p>
-					                    <strong><span style="font-family:;" new="" times="">A</span></strong>
+					                    <strong><span>A</span></strong>
 					                </p>
 					                <p>
-					                    <span style="font-family:;" new="" times=""><img width="300" src="https://static.igem.org/mediawiki/2014/4/48/Xmu_project_CircuitsDesign02.png"/></span>
+					                    <span><img width="300" src="https://static.igem.org/mediawiki/2014/4/48/Xmu_project_CircuitsDesign02.png"/></span>
 					                </p>
 					            </td>
 					            <td width="251" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p>
-					                    <strong><span style="font-family:;" new="" times="">B</span></strong>
+					                    <strong><span>B</span></strong>
 					                </p>
 					                <p>
-					                    <span style="font-family:;" new="" times=""><img width="300" src="https://static.igem.org/mediawiki/2014/4/48/Xmu_project_CircuitsDesign02.png"/></span>
+					                    <span><img width="300" src="https://static.igem.org/mediawiki/2014/4/48/Xmu_project_CircuitsDesign02.png"/></span>
 					                </p>
 					            </td>
@@ Line 618: / Line 696: @@
 					            <td width="553" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;" colspan="2">
 					                <p>
-					                    <strong><span style="font-family:;" new="" times="">Figure 2A</span></strong><span style="font-family:;" new="" times="">, Schematic of &nbsp; hyperbola. Point F1, F2 are the two focal points and Point A is on one of the &nbsp; hyperbola branches. The absolute value of the difference of the distance AF1, &nbsp; AF2 is equal to the constant k: |AF1-AF2|=k. <strong>2B</strong>, one branch of hyperbola can be defined by directrix and &nbsp; eccentricity. Point A is a spot on the curve, point F is the focus. AB is the &nbsp; distance between A and directrix. The eccentricity e equal to TF/AB, and &nbsp; e&gt;1.</span>
+					                    <strong><span>Figure 2A</span></strong><span>, Schematic of &nbsp; hyperbola. Point F1, F2 are the two focal points and Point A is on one of the &nbsp; hyperbola branches. The absolute value of the difference of the distance AF1, &nbsp; AF2 is equal to the constant k: |AF1-AF2|=k. <strong>2B</strong>, one branch of hyperbola can be defined by directrix and &nbsp; eccentricity. Point A is a spot on the curve, point F is the focus. AB is the &nbsp; distance between A and directrix. The eccentricity e equal to TF/AB, and &nbsp; e&gt;1.</span>
 					                </p>
 					            </td>
@@ Line 625: / Line 703: @@
 					</table>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<table cellspacing="0" cellpadding="0" width="600px" style="position:relative;margin-left:-300px;left:50%">
@@ Line 632: / Line 710: @@
 					            <td width="198" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p>
-					                    <span style="font-family:;" new="" times=""><img width="600" src="https://static.igem.org/mediawiki/2014/7/75/Xmu_project_CircuitsDesign04.png"/></span>
+					                    <span><img width="600" src="https://static.igem.org/mediawiki/2014/7/75/Xmu_project_CircuitsDesign04.png"/></span>
 					                </p>
 					            </td>
@@ Line 639: / Line 717: @@
 					            <td width="198" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p>
-					                    <strong><span style="font-family:;" new="" times="">Figure 2C</span></strong><span style="font-family:;" new="" times=""> Schematic of &nbsp; Parabola. Point A is a spot on the curve, point F is the focus. AB is the &nbsp; distance between A and directrix. The eccentricity e equals to AF/AB, and &nbsp; e=1.</span>
+					                    <strong><span>Figure 2C</span></strong><span> Schematic of &nbsp; Parabola. Point A is a spot on the curve, point F is the focus. AB is the &nbsp; distance between A and directrix. The eccentricity e equals to AF/AB, and &nbsp; e=1.</span>
 					                </p>
 					            </td>
@@ Line 646: / Line 724: @@
 					</table>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <strong><span style="font-family:;" new="" times="">Hypothesis </span></strong>
+					    <strong><span>Hypothesis </span></strong>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">We find that the focal points, the constant and the ratio are the key of conic section. Therefore, we can easily get any eclipse by presetting reasonable focal points with the acceptable constant, as well as parabola or hyperbola by a fixed ratio. Based on these, we put forward the following hypothesis:</span>
+					    <span>We find that the focal points, the constant and the ratio are the key of conic section. Therefore, we can easily get any eclipse by presetting reasonable focal points with the acceptable constant, as well as parabola or hyperbola by a fixed ratio. Based on these, we put forward the following hypothesis:</span>
 					</p>
 					<ol class=" list-paddingleft-2" style="list-style-type: decimal;">
 					    <li>
 					        <p style="color: rgb(0, 0, 0); font-size: 14px; font-style: normal; font-weight: normal; margin-top: 0px; margin-bottom: 0px;">
-					            <span style="font-family:;" new="" times="">If we spot stimulus on semi-solid plate, it will spread from the spotting center out to the periphery. In the spreading process, the concentration is negative correlation to the distance from center. And the concentration gradient of the stimulus will be formed and maintained for a long period.</span>
+					            <span>If we spot stimulus on semi-solid plate, it will spread from the spotting center out to the periphery. In the spreading process, the concentration is negative correlation to the distance from center. And the concentration gradient of the stimulus will be formed and maintained for a long period.</span>
 					        </p>
 					    </li>
 					    <li>
 					        <p style="color: rgb(0, 0, 0); font-family:;">
-					            <span style="font-family:;" new="" times="">There is a threshold ratio of the concentrations of inducer and repressor. This means that more repressor will cause more repression, hence more inducer is needed to relieve the repression, and vice versa.</span>
+					            <span>There is a threshold ratio of the concentrations of inducer and repressor. This means that more repressor will cause more repression, hence more inducer is needed to relieve the repression, and vice versa.</span>
 					        </p>
 					    </li>
 					</ol>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <strong><span style="font-family:;" new="" times="">Design of circuit</span></strong>
+					    <strong><span>Design of circuit</span></strong>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">Our circuit consists of two parts, of which one is named C (constraint), the other is named M (motile). (<strong>Figure 4</strong>):</span>
+					    <span>Our circuit consists of two parts, of which one is named C (constraint), the other is named M (motile). (<strong>Figure 4</strong>):</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<ol class=" list-paddingleft-2" style="list-style-type: decimal;">
 					    <li>
 					        <p style="color: rgb(0, 0, 0); font-size: 14px; font-style: normal; font-weight: normal; margin-top: 0px; margin-bottom: 0px;">
-					            <span style="font-family:;" new="" times="">We build our circuit in <em>E.coli</em> <em>CL-1</em> which lacks gene <em>lacI</em> and <em>CheZ </em>(</span><span style="font-family:;" new="" times="">Δ</span><span style="font-family:;" new="" times="">lacI, </span><span style="font-family:;" new="" times="">Δ</span><span style="font-family:;" new="" times="">CheZ). At the absence of CheZ,<em> CL-1</em> adopts non-motile <a name="OLE_LINK2"></a>phenotype.</span>
+					            <span>We build our circuit in <em>E.coli</em> <em>CL-1</em> which lacks gene <em>lacI</em> and <em>CheZ </em>(</span><span>Δ</span><span>lacI, </span><span>Δ</span><span>CheZ). At the absence of CheZ,<em> CL-1</em> adopts non-motile <a name="OLE_LINK2"></a>phenotype.</span>
 					        </p>
 					    </li>
 					    <li>
 					        <p style="color: rgb(0, 0, 0); font-family:;">
-					            <span style="font-family:;" new="" times="">Without any exogenous stimulus, <em>E.coli</em> will produce background amount of AraC to repress pBAD in limit degree. Even when L-arabinose isn’t involved in, promoter pBAD has expression leakage, so that part <strong>C </strong>will produces protein LacI which can bind to the operon of promoter pLac and thus repress its transcription. Because L-arabinose could induce pBAD, within certain concentration range, more L-arabinose involved in means that part <strong>C</strong> could produce more LacI to repress the expression of pLac. Because of its ability to constrain chemotaxis, this part is named C (abbreviate from constraint).</span>
+					            <span>Without any exogenous stimulus, <em>E.coli</em> will produce background amount of AraC to repress pBAD in limit degree. Even when L-arabinose isn’t involved in, promoter pBAD has expression leakage, so that part <strong>C </strong>will produces protein LacI which can bind to the operon of promoter pLac and thus repress its transcription. Because L-arabinose could induce pBAD, within certain concentration range, more L-arabinose involved in means that part <strong>C</strong> could produce more LacI to repress the expression of pLac. Because of its ability to constrain chemotaxis, this part is named C (abbreviate from constraint).</span>
 					        </p>
 					    </li>
 					    <li>
 					        <p style="color: rgb(0, 0, 0); font-family:;">
-					            <span style="font-family:;" new="" times="">When IPTG is involved in, it can relieve the repression from LacI, therefore protein CheZ is produced to make our engineering bacteria (<em>CL-1</em>) regain motile ability. Within certain L-Arabinose concentration range which means certain constraint condition, more cheZ is produced with more IPTG involved in leading to stronger motile ability. Because of its ability to make <em>CL-1</em> motile, this part is named <strong>M</strong> (abbreviate from motile).</span>
+					            <span>When IPTG is involved in, it can relieve the repression from LacI, therefore protein CheZ is produced to make our engineering bacteria (<em>CL-1</em>) regain motile ability. Within certain L-Arabinose concentration range which means certain constraint condition, more cheZ is produced with more IPTG involved in leading to stronger motile ability. Because of its ability to make <em>CL-1</em> motile, this part is named <strong>M</strong> (abbreviate from motile).</span>
 					        </p>
 					    </li>
@@ Line 709: / Line 787: @@
 					            <td width="321" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p>
-					                    <strong><span style="font-family:;" new="" times=""><img width="600" src="https://static.igem.org/mediawiki/2014/4/45/Xmu_project_CircuitsDesign05.png"/></span></strong>
+					                    <strong><span><img width="600" src="https://static.igem.org/mediawiki/2014/4/45/Xmu_project_CircuitsDesign05.png"/></span></strong>
 					                </p>
 					            </td>
@@ Line 716: / Line 794: @@
 					            <td width="321" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p style="margin: 0px 0px 0px 28px; text-align: left;">
-					                    <strong><span style="font-family:;" new="" times="">Figure 4 </span></strong><span style="font-family:;" new="" times="">Part<strong> C </strong>produces LacI to repress the &nbsp; expression of part <strong>M</strong>. Part <strong>M</strong> could produce CheZ to make <em>CL-1 </em>regain motile ability.</span>
+					                    <strong><span>Figure 4 </span></strong><span>Part<strong> C </strong>produces LacI to repress the &nbsp; expression of part <strong>M</strong>. Part <strong>M</strong> could produce CheZ to make <em>CL-1 </em>regain motile ability.</span>
 					                </p>
 					            </td>
@@ Line 723: / Line 801: @@
 					</table>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
@@ Line 831: / Line 909: @@
 					</table>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <strong><span style="font-family:;" new="" times="">Eclipse</span></strong>
+					    <strong><span>Eclipse</span></strong>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">If we spot IPTG on the center of the semi-solid culture medium, concentration gradient will be formed as <strong>Figure</strong> <strong>3A</strong>. Larger circle represents lower concentration with a lower number labeled.</span>
+					    <span>If we spot IPTG on the center of the semi-solid culture medium, concentration gradient will be formed as <strong>Figure</strong> <strong>3A</strong>. Larger circle represents lower concentration with a lower number labeled.</span>
 					</p>
 					<table width="567" cellspacing="0" cellpadding="0" width="600px" style="position:relative;margin-left:-300px;left:50%">
@@ Line 850: / Line 928: @@
 					            <td width="222" height="279" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p>
-					                    <strong><span style="font-family:;" new="" times="">A</span></strong>
+					                    <strong><span>A</span></strong>
 					                </p>
 					                <p style="text-align: left;">
-					                    <span style="font-family:;" new="" times=""><img width="300" src="https://static.igem.org/mediawiki/2014/d/d4/Xmu_project_CircuitsDesign09.png"/></span>
+					                    <span><img width="300" src="https://static.igem.org/mediawiki/2014/d/d4/Xmu_project_CircuitsDesign09.png"/></span>
 					                </p>
 					            </td>
 					            <td width="344" height="279" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p>
-					                    <strong><span style="font-family:;" new="" times="">B</span></strong>
+					                    <strong><span>B</span></strong>
 					                </p>
 					                <p>
-					                    <span style="font-family:;" new="" times=""><img width="300" src="https://static.igem.org/mediawiki/2014/1/16/Xmu_project_CircuitsDesign10.png"/></span>
+					                    <span><img width="300" src="https://static.igem.org/mediawiki/2014/1/16/Xmu_project_CircuitsDesign10.png"/></span>
 					                </p>
 					            </td>
@@ Line 868: / Line 946: @@
 					            <td width="567" height="21" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;" colspan="2">
 					                <p>
-					                    <strong><span style="font-family:;" new="" times="">Figure</span></strong><span style="font-family:;" new="" times=""> <strong>3</strong> Schematic of concentration &nbsp; gradient. <strong>A,</strong> single point &nbsp; spreading model. Larger number represents higher concentration while smaller &nbsp; number represents lower concentration. <strong>B</strong>, &nbsp; double points spreading model. Letter A~J represent equal concentration &nbsp; points on the ellipse with two labeled number added up to 6. Red curve &nbsp; represents the ellipse with two focal points on the IPTG spots.</span>
+					                    <strong><span>Figure</span></strong><span> <strong>3</strong> Schematic of concentration &nbsp; gradient. <strong>A,</strong> single point &nbsp; spreading model. Larger number represents higher concentration while smaller &nbsp; number represents lower concentration. <strong>B</strong>, &nbsp; double points spreading model. Letter A~J represent equal concentration &nbsp; points on the ellipse with two labeled number added up to 6. Red curve &nbsp; represents the ellipse with two focal points on the IPTG spots.</span>
 					                </p>
 					            </td>
@@ Line 875: / Line 953: @@
 					</table>
 					<p>
-					    <span style="font-family:;" new="" times="">If we spot two IPTG simultaneously, two spots’ concentration gradient will interact with each other to form the ellipse boundary as image in <strong>Figure 3B</strong>. </span>
+					    <span>If we spot two IPTG simultaneously, two spots’ concentration gradient will interact with each other to form the ellipse boundary as image in <strong>Figure 3B</strong>. </span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">If we spot <em>E.coli</em> <em>CL-1</em> with circuit above transformed in at the center between two IPTG spots on semi-solid culture plate as the image in <strong>Figure 5</strong>. The concentration of L-Arabinose added in the culture medium determines the degree of constraint from part <strong>C</strong>. Each red ellipse curve represents equal IPTG concentration, and there are a series of such ellipses from inside to outside represent IPTG concentrations from highest to lowest. One of the ellipses is the <strong>critical line</strong> indicating that the constraint from part <strong>C</strong> can just be relieved by the certain concentration of IPTG. Initially, as the concentration of IPTG is enough to relieve the constraint, bacteria can swim from the center out to periphery. When the bacteria swim out of the <strong>critical line</strong>, the concentration of IPTG can’t relieve the constraint, so the bacteria adopt non-motile phenotype. On the contrary, when the bacteria are inside of the critical line, they adopt motile phenotype. When the bacteria swim from inside to outside, motile bacteria become non-motile, so the bacteria will aggregate outside the <strong>critical line</strong> while</span><span style="font-family:;" new="" times=""> the b</span><span style="font-family:;" new="" times="">acteria density inside the<strong> critical line</strong> will decline. Thus, an ellipse boundary is formed.</span>
+					    <span>If we spot <em>E.coli</em> <em>CL-1</em> with circuit above transformed in at the center between two IPTG spots on semi-solid culture plate as the image in <strong>Figure 5</strong>. The concentration of L-Arabinose added in the culture medium determines the degree of constraint from part <strong>C</strong>. Each red ellipse curve represents equal IPTG concentration, and there are a series of such ellipses from inside to outside represent IPTG concentrations from highest to lowest. One of the ellipses is the <strong>critical line</strong> indicating that the constraint from part <strong>C</strong> can just be relieved by the certain concentration of IPTG. Initially, as the concentration of IPTG is enough to relieve the constraint, bacteria can swim from the center out to periphery. When the bacteria swim out of the <strong>critical line</strong>, the concentration of IPTG can’t relieve the constraint, so the bacteria adopt non-motile phenotype. On the contrary, when the bacteria are inside of the critical line, they adopt motile phenotype. When the bacteria swim from inside to outside, motile bacteria become non-motile, so the bacteria will aggregate outside the <strong>critical line</strong> while</span><span> the b</span><span>acteria density inside the<strong> critical line</strong> will decline. Thus, an ellipse boundary is formed.</span>
 					</p>
 					<table cellspacing="0" cellpadding="0" width="600px" style="position:relative;margin-left:-300px;left:50%">
@@ Line 888: / Line 966: @@
 					            <td width="425" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p>
-					                    <span style="font-family:;" new="" times=""><img width="600" src="https://static.igem.org/mediawiki/2014/1/14/Xmu_project_CircuitsDesign11.png"/></span>
+					                    <span><img width="600" src="https://static.igem.org/mediawiki/2014/1/14/Xmu_project_CircuitsDesign11.png"/></span>
 					                </p>
 					            </td>
@@ Line 895: / Line 973: @@
 					            <td width="425" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p>
-					                    <strong><span style="font-family:;" new="" times="">Figure 5</span></strong><span style="font-family:;" new="" times=""> Schematic of &nbsp; Critical Line model for ellipse. Critical line represents the IPTG &nbsp; concentration which can just relieve the repression from part <strong>C</strong>. So that <em>CL-1</em> is motile inside the ellipse while non-motile outside the &nbsp; ellipse.</span>
+					                    <strong><span>Figure 5</span></strong><span> Schematic of &nbsp; Critical Line model for ellipse. Critical line represents the IPTG &nbsp; concentration which can just relieve the repression from part <strong>C</strong>. So that <em>CL-1</em> is motile inside the ellipse while non-motile outside the &nbsp; ellipse.</span>
 					                </p>
 					            </td>
@@ Line 902: / Line 980: @@
 					</table>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <strong><span style="font-family:;" new="" times="">Parabola and hyperbola</span></strong>
+					    <strong><span>Parabola and hyperbola</span></strong>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">We got the optimum concentrations of IPTG and L-arabinose as inducer and repressor through preliminary experiments. Because the concentration of stimulus will decrease during spreading, so does their effect, we use IPTG and L-arabinose of which concentrations are a little bit higher than the optimum values for our experiments.</span>
+					    <span>We got the optimum concentrations of IPTG and L-arabinose as inducer and repressor through preliminary experiments. Because the concentration of stimulus will decrease during spreading, so does their effect, we use IPTG and L-arabinose of which concentrations are a little bit higher than the optimum values for our experiments.</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">We draw a straight line with L-arabinose on the semi-solid culture medium, and a spot with the mixture of IPTG and CL-1 on one side of the line. In the area around the spot, the induction of IPTG is stronger than the repression of L-arabinose, <em>CheZ</em> is expressed and the bacteria adopt motile phenotype. However, when they approach the line where the repressor have a greater effect on the motility, they will lost their motile phenotype and stop. </span>
+					    <span>We draw a straight line with L-arabinose on the semi-solid culture medium, and a spot with the mixture of IPTG and CL-1 on one side of the line. In the area around the spot, the induction of IPTG is stronger than the repression of L-arabinose, <em>CheZ</em> is expressed and the bacteria adopt motile phenotype. However, when they approach the line where the repressor have a greater effect on the motility, they will lost their motile phenotype and stop. </span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">According to the hypothesis 2, on the threshold ratio of the concentration of inducer and repressor, their effects are offset and a critical line is formed. Distances of the points on the critical line to the IPTG spot (focus), and the L-arabinose line (directrix) are in a fixed ratio (eccentricity). If the ratio is equal to 1, the critical line is a parabola. If the ratio is larger than 1, it is a branch of a hyperbola.</span>
+					    <span>According to the hypothesis 2, on the threshold ratio of the concentration of inducer and repressor, their effects are offset and a critical line is formed. Distances of the points on the critical line to the IPTG spot (focus), and the L-arabinose line (directrix) are in a fixed ratio (eccentricity). If the ratio is equal to 1, the critical line is a parabola. If the ratio is larger than 1, it is a branch of a hyperbola.</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<table cellspacing="0" cellpadding="0" width="600px" style="position:relative;margin-left:-300px;left:50%">
@@ Line 936: / Line 1,014: @@
 					            <td width="315" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p>
-					                    <span style="font-family:;" new="" times=""><img width="600" src="https://static.igem.org/mediawiki/2014/1/14/Xmu_project_CircuitsDesign12.png"/></span>
+					                    <span><img width="600" src="https://static.igem.org/mediawiki/2014/1/14/Xmu_project_CircuitsDesign12.png"/></span>
 					                </p>
 					            </td>
@@ Line 943: / Line 1,021: @@
 					            <td width="315" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p>
-					                    <strong><span style="font-family:;" new="" times="">Figure 6 </span></strong><span style="font-family:;" new="" times="">Schematic of &nbsp; Critical Line model for parabola and hyperbola. CL-1 becomes no-motile on the &nbsp; left side of the critical line while motile on the other side. The &nbsp; eccentricity e equals to PF/PB. If e=1, we define the critical line as &nbsp; parabola. If e&gt;1, we define that as one branch of hyperbola.</span>
+					                    <strong><span>Figure 6 </span></strong><span>Schematic of &nbsp; Critical Line model for parabola and hyperbola. CL-1 becomes no-motile on the &nbsp; left side of the critical line while motile on the other side. The &nbsp; eccentricity e equals to PF/PB. If e=1, we define the critical line as &nbsp; parabola. If e&gt;1, we define that as one branch of hyperbola.</span>
 					                </p>
 					            </td>
@@ Line 950: / Line 1,028: @@
 					</table>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <strong><span style="font-family:;" new="" times="">Other function curves</span></strong>
+					    <strong><span>Other function curves</span></strong>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">After explorations, we found a new way to spot bacteria and stimulus and got a special function curves.</span>
+					    <span>After explorations, we found a new way to spot bacteria and stimulus and got a special function curves.</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">We draw two spots on the semi-solid culture medium, one with L-arabinose (Figure 7A) and the other with the mixture of IPTG and CL-1. Similarly, on the threshold ratio of the concentration of inducer and repressor, their effects are offset and a<strong> critical line</strong> is formed. Distances of the points on the <strong>critical line</strong> to the spot A and the spot B are in a fixed ratio (Figure 7B). Actually, as the critical line is quiet similar to hyperbola, we name it quasi-hyperbola.</span>
+					    <span>We draw two spots on the semi-solid culture medium, one with L-arabinose (Figure 7A) and the other with the mixture of IPTG and CL-1. Similarly, on the threshold ratio of the concentration of inducer and repressor, their effects are offset and a<strong> critical line</strong> is formed. Distances of the points on the <strong>critical line</strong> to the spot A and the spot B are in a fixed ratio (Figure 7B). Actually, as the critical line is quiet similar to hyperbola, we name it quasi-hyperbola.</span>
 					</p>
 					<table cellspacing="0" cellpadding="0" width="600px" style="position:relative;margin-left:-300px;left:50%">
@@ Line 969: / Line 1,047: @@
 					            <td width="252" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p>
-					                    <span style="font-family:;" new="" times="">A</span>
+					                    <span>A</span>
 					                </p>
 					                <p>
-					                    <span style="font-family:;" new="" times=""><img width="270" src="https://static.igem.org/mediawiki/2014/5/53/Xmu_project_CircuitsDesign13.png"/></span>
+					                    <span><img width="270" src="https://static.igem.org/mediawiki/2014/5/53/Xmu_project_CircuitsDesign13.png"/></span>
 					                </p>
 					            </td>
 					            <td width="296" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;">
 					                <p>
-					                    <span style="font-family:;" new="" times="">B</span>
+					                    <span>B</span>
 					                </p>
 					                <p>
-					                    <span style="font-family:;" new="" times=""><img width="330" src="https://static.igem.org/mediawiki/2014/0/04/Xmu_project_CircuitsDesign14.png"/></span>
+					                    <span><img width="330" src="https://static.igem.org/mediawiki/2014/0/04/Xmu_project_CircuitsDesign14.png"/></span>
 					                </p>
 					            </td>
@@ Line 987: / Line 1,065: @@
 					            <td width="548" valign="top" style="padding: 0px 7px; border: rgb(0, 0, 0); border-image: none; background-color: transparent;" colspan="2">
 					                <p style="margin: 0px 0px 0px 7px;">
-					                    <strong><span style="font-family:;" new="" times="">Figure 7A</span></strong><span style="font-family:;" new="" times=""> Schematic of &nbsp; quasi-hyperbola formation on semi-solid medium culture. <strong>7B</strong> Actual experiment result is shown.</span>
+					                    <strong><span>Figure 7A</span></strong><span> Schematic of &nbsp; quasi-hyperbola formation on semi-solid medium culture. <strong>7B</strong> Actual experiment result is shown.</span>
 					                </p>
 					            </td>
@@ Line 994: / Line 1,072: @@
 					</table>
 					<p>
-					    <span style="font-family:;" new="" times="">&nbsp;</span>
+					    <span>&nbsp;</span>
 					</p>
 					<p>
-					    <span style="font-family:;" new="" times="">Other interesting ways to spot bacteria and stimulus are waiting to be discovered, and the idea can be extended to other function curves and patterns.</span>
+					    <span>Other interesting ways to spot bacteria and stimulus are waiting to be discovered, and the idea can be extended to other function curves and patterns.</span>
 					</p>
 					<p>
@@ Line 1,008: / Line 1,086: @@
 				<img  id="Project_Black_hole_title" class="Project_title" src="https://static.igem.org/mediawiki/2014/a/a9/Xmu_project_background.png" />
 				<div id="Project_Black_hole_wd">
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;">T</span><span style="font-family: Times New Roman;">housands of years ago in C</span><span style="font-family: Times New Roman;">hina, people </span><span style="font-family: Times New Roman;">began to preserve food by curing them</span><span style="font-family: Times New Roman;"> which was recorded in</span><span style="font-family: Times New Roman; font-style: italic;"> Qimin Yaoshu</span><span style="font-family: Times New Roman;"> around 540 AD</span><span style="font-family: Times New Roman;"> (</span><span style="font-family: Times New Roman; font-weight: 700;">Figure 1</span><span style="font-family: Times New Roman;">)</span><span style="font-family: Times New Roman;">. Curing is any of various food preservation and flavoring processes of foods such as meat, fish and vegetables, by the addition of</span><span style="font-family: Times New Roman;"> a combination of salt, nitrate</span><span style="font-family: Times New Roman;">s</span><span style="font-family: Times New Roman;">, nitrite or sugar and it is one of the oldest m</span><span style="font-family: Times New Roman;">ethods of preserving food. T</span><span style="font-family: Times New Roman;">able salt is the primary ingredient used in food curing. Removal of water and addition of salt to meat creates a solute-rich environment where osmotic pressure draws water out of microorganisms, slowing down their growth. Doing this requires a concentration of salt of nearly 20%. </span><span style="font-family: Times New Roman;">It has already been proved that 5% concentration of NaCl could inhibit the growth of </span><span style="font-family: Times New Roman; font-style: italic;">E.coli</span><span style="font-family: Times New Roman;">. However, utilizing </span><span style="font-family: Times New Roman;">hyperosmotic pressure</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">to kill </span><span style="font-family: Times New Roman; font-style: italic;">E.coli</span>
+					    <span>T</span><span>housands of years ago in C</span><span>hina, people </span><span>began to preserve food by curing them</span><span> which was recorded in</span><span style="font-family: ; font-style: italic;"> Qimin Yaoshu</span><span> around 540 AD</span><span> (</span><span style="font-family: ; font-weight: 700;">Figure 1</span><span>)</span><span>. Curing is any of various food preservation and flavoring processes of foods such as meat, fish and vegetables, by the addition of</span><span> a combination of salt, nitrate</span><span>s</span><span>, nitrite or sugar and it is one of the oldest m</span><span>ethods of preserving food. T</span><span>able salt is the primary ingredient used in food curing. Removal of water and addition of salt to meat creates a solute-rich environment where osmotic pressure draws water out of microorganisms, slowing down their growth. Doing this requires a concentration of salt of nearly 20%. </span><span>It has already been proved that 5% concentration of NaCl could inhibit the growth of </span><span style="font-family: ; font-style: italic;">E.coli</span><span>. However, utilizing </span><span>hyperosmotic pressure</span><span> </span><span>to kill </span><span style="font-family: ; font-style: italic;">E.coli</span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;"> haven’t been fully explored</span><span style="font-family: Times New Roman;"> in synthetic biology</span><span style="font-family: Times New Roman;">. T</span><span style="font-family: Times New Roman;">his year, our team have put efforts on this </span><span style="font-family: Times New Roman;">topic </span><span style="font-family: Times New Roman;">and develop</span><span style="font-family: Times New Roman;">ed</span><span style="font-family: Times New Roman;"> a system </span><span style="font-family: Times New Roman;">that</span><span style="font-family: Times New Roman;"> will contribute to biosafety.</span>
+					    <span> haven’t been fully explored</span><span> in synthetic biology</span><span>. T</span><span>his year, our team have put efforts on this </span><span>topic </span><span>and develop</span><span>ed</span><span> a system </span><span>that</span><span> will contribute to biosafety.</span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
 					    &nbsp;
 					</p>
@@ Line 1,021: / Line 1,099: @@
 					        <tr class="firstRow">
 					            <td style="padding: 0px 7px; border: 1px rgb(0, 0, 0); width: 553px; vertical-align: top;">
-					                <p style="font-family: Times New Roman;">
+					                <p>
 					                    <img width="540" src="https://static.igem.org/mediawiki/2014/5/59/Xmu_project_BlackHole01.png"/>
 					                </p>
@@ Line 1,028: / Line 1,106: @@
 					        <tr>
 					            <td style="border-width: medium 1px 1px; border-color: currentColor rgb(0, 0, 0) rgb(0, 0, 0); padding: 0px 7px; width: 553px; vertical-align: top;">
-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <span style="font-family: Times New Roman; font-weight: 700;">Figure 1</span><span style="font-family: Times New Roman;"> The production process of curing food.</span>
+					                    <span style="font-family: ; font-weight: 700;">Figure 1</span><span> The production process of curing food.</span>
 					                </p>
 					            </td>
@@ Line 1,038: / Line 1,116: @@
 					    </tbody>
 					</table>
-					<p style="font-family: Times New Roman;">
+					<p>
 					    &nbsp;
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
 					    &nbsp;
 					</p>
@@ Line 1,048: / Line 1,126: @@
 					        <tr class="firstRow">
 					            <td style="padding: 0px 7px; border: currentColor; width: 553px; vertical-align: top;">
-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <img width="539" height="302" style="font-family: Times New Roman;" src="https://static.igem.org/mediawiki/2014/1/1f/Xmu_project_BlackHole02.png"/>
+					                    <img width="539" height="302" src="https://static.igem.org/mediawiki/2014/1/1f/Xmu_project_BlackHole02.png"/>
 					                </p>
 					            </td>
@@ Line 1,055: / Line 1,133: @@
 					        <tr>
 					            <td style="padding: 0px 7px; border: currentColor; width: 553px; vertical-align: top;">
-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <span style="font-family: Times New Roman; font-weight: 700;">Figure 2</span><span style="font-family: Times New Roman;"> The </span><span style="font-family: Times New Roman;">schematic of osmotic-taxis design.</span>
+					                    <span style="font-family: ; font-weight: 700;">Figure 2</span><span> The </span><span>schematic of osmotic-taxis design.</span>
 					                </p>
 					            </td>
@@ Line 1,065: / Line 1,143: @@
 					    </tbody>
 					</table>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;"> </span>
+					    <span> </span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman; font-style: italic;">E</span><span style="font-family: Times New Roman; font-style: italic;">.c</span><span style="font-family: Times New Roman; font-style: italic;">oli</span><span style="font-family: Times New Roman;"> make</span><span style="font-family: Times New Roman;">s</span><span style="font-family: Times New Roman;"> use of the EnvZ/OmpR system to mediate signal transduction in response to environmental osmolarity changes. EnvZ, a histidine kinase, undergoes trans-autophosphorylation, then the </span><span style="font-family: Times New Roman;">high-</span><span style="font-family: Times New Roman;">energy phosphoryl group is subsequently transferred to OmpR, a response regulator</span><span style="font-family: Times New Roman;">.</span>
+					    <span style="font-family: ; font-style: italic;">E</span><span style="font-family: ; font-style: italic;">.c</span><span style="font-family: ; font-style: italic;">oli</span><span> make</span><span>s</span><span> use of the EnvZ/OmpR system to mediate signal transduction in response to environmental osmolarity changes. EnvZ, a histidine kinase, undergoes trans-autophosphorylation, then the </span><span>high-</span><span>energy phosphoryl group is subsequently transferred to OmpR, a response regulator</span><span>.</span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">In </span><span style="font-family: Times New Roman;">our</span><span style="font-family: Times New Roman;"> system, we </span><span style="font-family: Times New Roman;">involved OmpR-controlled promoter</span><span style="font-family: Times New Roman;"> (PompC)</span><span style="font-family: Times New Roman;"> in</span><span style="font-family: Times New Roman;"> (Figure 2)</span><span style="font-family: Times New Roman;">. </span><span style="font-family: Times New Roman;">The expression strength of PompC</span><span style="font-family: Times New Roman;"> is </span><span style="font-family: Times New Roman;">depending upon the medium osmolarity. </span><span style="font-family: Times New Roman;">When medium osmolarity is increasing, the EnvZ will phosphorylate more OmpR into phosphorylated OmpR (OmpR-P), </span><span style="font-family: Times New Roman;">resulting</span><span style="font-family: Times New Roman;"> in stronger expression strength of PompR.</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">In our circuitry design, </span><span style="font-family: Times New Roman; font-style: italic;">CheZ</span><span style="font-family: Times New Roman;"> is upstream regulated by PompR.</span>
+					    <span> </span><span>In </span><span>our</span><span> system, we </span><span>involved OmpR-controlled promoter</span><span> (PompC)</span><span> in</span><span> (Figure 2)</span><span>. </span><span>The expression strength of PompC</span><span> is </span><span>depending upon the medium osmolarity. </span><span>When medium osmolarity is increasing, the EnvZ will phosphorylate more OmpR into phosphorylated OmpR (OmpR-P), </span><span>resulting</span><span> in stronger expression strength of PompR.</span><span> </span><span>In our circuitry design, </span><span style="font-family: ; font-style: italic;">CheZ</span><span> is upstream regulated by PompR.</span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
 					    &nbsp;
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;">We use semi-solid medium culture with gradient concentration of sucrose</span><span style="font-family: Times New Roman;"> to characterize the device (BBa_K1412008)</span><span style="font-family: Times New Roman;">.</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">And we assume that </span><span style="font-family: Times New Roman;">the motile ability is proportional to the moving radius.</span><span style="font-family: Times New Roman;"> In t</span><span style="font-family: Times New Roman;">he plot (Figure 3), when </span><span style="font-family: Times New Roman;">no sucrose added in, the motile ability is the weakest. The motile ability keeps growing </span><span style="font-family: Times New Roman;">as </span><span style="font-family: Times New Roman;">the concentration of sucrose increase</span><span style="font-family: Times New Roman;">s </span><span style="font-family: Times New Roman;">from 0 to 4%. Then t</span><span style="font-family: Times New Roman;">he motile ability goes </span><span style="font-family: Times New Roman;">down slightly as the sucrose concentration increased from 4% to 10%, but </span><span style="font-family: Times New Roman;">the ability is</span><span style="font-family: Times New Roman;"> still stronger than that at</span><span style="font-family: Times New Roman;"> concentration 0. We can draw a </span><span style="font-family: Times New Roman;">conclusion that our device is working </span><span style="font-family: Times New Roman;">as expectation</span><span style="font-family: Times New Roman;">, the motile ability </span><span style="font-family: Times New Roman;">goes</span><span style="font-family: Times New Roman;"> down</span><span style="font-family: Times New Roman;"> (4%~10%)</span><span style="font-family: Times New Roman;"> because of the inhibition from hyperosmotic pressure.</span>
+					    <span>We use semi-solid medium culture with gradient concentration of sucrose</span><span> to characterize the device (BBa_K1412008)</span><span>.</span><span> </span><span>And we assume that </span><span>the motile ability is proportional to the moving radius.</span><span> In t</span><span>he plot (Figure 3), when </span><span>no sucrose added in, the motile ability is the weakest. The motile ability keeps growing </span><span>as </span><span>the concentration of sucrose increase</span><span>s </span><span>from 0 to 4%. Then t</span><span>he motile ability goes </span><span>down slightly as the sucrose concentration increased from 4% to 10%, but </span><span>the ability is</span><span> still stronger than that at</span><span> concentration 0. We can draw a </span><span>conclusion that our device is working </span><span>as expectation</span><span>, the motile ability </span><span>goes</span><span> down</span><span> (4%~10%)</span><span> because of the inhibition from hyperosmotic pressure.</span>
 					</p>
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-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <img width="537" height="470" style="font-family: Times New Roman;" src="https://static.igem.org/mediawiki/2014/a/ab/Xmu_project_BlackHole03.png"/>
+					                    <img width="537" height="470" src="https://static.igem.org/mediawiki/2014/a/ab/Xmu_project_BlackHole03.png"/>
 					                </p>
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-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <span style="font-family: Times New Roman; font-weight: 700;">Figure 3</span><span style="font-family: Times New Roman;"> The plot of moving radius versus sucrose concentration. The four curves were measured after 10h, 11h, 12h and 16.5h respectively.</span>
+					                    <span style="font-family: ; font-weight: 700;">Figure 3</span><span> The plot of moving radius versus sucrose concentration. The four curves were measured after 10h, 11h, 12h and 16.5h respectively.</span>
 					                </p>
 					            </td>
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 					    </tbody>
 					</table>
-					<p style="font-family: Times New Roman;">
+					<p>
 					    &nbsp;
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;">Based on </span><span style="font-family: Times New Roman;">the</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">characterization, we </span><span style="font-family: Times New Roman;">spotted </span><span style="font-family: Times New Roman;">hyperosmotic pressure spot and reprogrammed</span><span style="font-family: Times New Roman; font-style: italic;"> CL-1</span><span style="font-family: Times New Roman;"> spot </span><span style="font-family: Times New Roman;">on semi-solid medium culture </span><span style="font-family: Times New Roman;">as Figure 4 show</span><span style="font-family: Times New Roman;">s</span><span style="font-family: Times New Roman;">.</span><span style="font-family: Times New Roman;"> The concentration will decrease</span><span style="font-size: 14px;"> </span><span style="font-family: Times New Roman;">w</span><span style="font-family: Times New Roman;">ith the increase of the distance away from hyperosmotic pressure spot</span><span style="font-family: Times New Roman;">. As the osmotic pressure is </span><span style="font-family: Times New Roman;">proportional to the medium concentration. The moving tendency of reprogrammed </span><span style="font-family: Times New Roman; font-style: italic;">CL-1</span><span style="font-family: Times New Roman;"> will orient to the hyperosmotic pressure spot.</span><span style="font-family: Times New Roman;"> Even at the inhibiting osmotic pressure, the motile ability is still stronger than that without </span><span style="font-family: Times New Roman;">any inducer.</span><span style="font-family: Times New Roman;"> So reprogrammed </span><span style="font-family: Times New Roman; font-style: italic;">CL-1 </span><span style="font-family: Times New Roman;">may even swim </span><span style="font-family: Times New Roman;">t</span><span style="font-family: Times New Roman;">owards the high-osmotic site and die</span><span style="font-family: Times New Roman;">. The killing mechan</span><span style="font-family: Times New Roman;">ism is just like the black hole. W</span><span style="font-family: Times New Roman;">hen the bacteria </span><span style="font-family: Times New Roman;">move</span><span style="font-family: Times New Roman;"> into the “event horizon” </span><span style="font-family: Times New Roman;">w</span><span style="font-family: Times New Roman;">here the osmotic pressure reaches to the critical value named the killing osmotic pressure</span><span style="font-family: Times New Roman;">, the bacteria can’t go out of the border</span>
+					    <span>Based on </span><span>the</span><span> </span><span>characterization, we </span><span>spotted </span><span>hyperosmotic pressure spot and reprogrammed</span><span style="font-family: ; font-style: italic;"> CL-1</span><span> spot </span><span>on semi-solid medium culture </span><span>as Figure 4 show</span><span>s</span><span>.</span><span> The concentration will decrease</span><span style="font-size: 14px;"> </span><span>w</span><span>ith the increase of the distance away from hyperosmotic pressure spot</span><span>. As the osmotic pressure is </span><span>proportional to the medium concentration. The moving tendency of reprogrammed </span><span style="font-family: ; font-style: italic;">CL-1</span><span> will orient to the hyperosmotic pressure spot.</span><span> Even at the inhibiting osmotic pressure, the motile ability is still stronger than that without </span><span>any inducer.</span><span> So reprogrammed </span><span style="font-family: ; font-style: italic;">CL-1 </span><span>may even swim </span><span>t</span><span>owards the high-osmotic site and die</span><span>. The killing mechan</span><span>ism is just like the black hole. W</span><span>hen the bacteria </span><span>move</span><span> into the “event horizon” </span><span>w</span><span>here the osmotic pressure reaches to the critical value named the killing osmotic pressure</span><span>, the bacteria can’t go out of the border</span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;"> and be killed finally.</span>
+					    <span> and be killed finally.</span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
 					    &nbsp;
 					</p>
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-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <img width="500" height="500" style="font-family: Times New Roman;" src="https://static.igem.org/mediawiki/2014/f/f0/Xmu_project_BlackHole04.png"/>
+					                    <img width="500" height="500" src="https://static.igem.org/mediawiki/2014/f/f0/Xmu_project_BlackHole04.png"/>
 					                </p>
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-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <img width="500" style="font-family: Times New Roman;" src="https://static.igem.org/mediawiki/2014/d/dd/Xmu_project_BlackHole05.png"/>
+					                    <img width="500" src="https://static.igem.org/mediawiki/2014/d/dd/Xmu_project_BlackHole05.png"/>
 					                </p>
 					            </td>
@@ Line 1,141: / Line 1,219: @@
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-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <span style="font-family: Times New Roman; font-weight: 700;">Figure </span><span style="font-family: Times New Roman; font-weight: 700;">4</span><span style="font-family: Times New Roman;"> Schematic of k</span><span style="font-family: Times New Roman;">illing bacteria by black hole.</span>
+					                    <span style="font-family: ; font-weight: 700;">Figure </span><span style="font-family: ; font-weight: 700;">4</span><span> Schematic of k</span><span>illing bacteria by black hole.</span>
 					                </p>
 					            </td>
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 					    </tbody>
 					</table>
-					<p style="font-family: Times New Roman;">
+					<p>
 					    &nbsp;
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;">It’s very cheap and </span><span style="font-family: Times New Roman;">accessible</span><span style="font-family: Times New Roman;"> to get the source (such as NaCl and sucrose) to create </span><span style="font-family: Times New Roman;">hyperosmotic pressure</span><span style="font-family: Times New Roman;">, w</span><span style="font-family: Times New Roman;">hile </span><span style="font-family: Times New Roman;">antibiotics</span><span style="font-family: Times New Roman;"> is expensive and have </span><span style="font-family: Times New Roman;">a bad</span><span style="font-family: Times New Roman;"> effect</span><span style="font-family: Times New Roman;"> on </span><span style="font-family: Times New Roman;">environmental microbiology</span><span style="font-family: Times New Roman;"> because of drug resistance. However, the source to </span><span style="font-family: Times New Roman;">hyperosmotic pressure</span><span style="font-family: Times New Roman;"> is </span><span style="font-family: Times New Roman;">environmentally</span><span style="font-family: Times New Roman;"> friendly and won’t generate the risk of drug resistance. If our black hole system could be fully developed, it will </span><span style="font-family: Times New Roman;">reduce the access barriers to microbiology research especially for the scientists from poor countries.</span>
+					    <span>It’s very cheap and </span><span>accessible</span><span> to get the source (such as NaCl and sucrose) to create </span><span>hyperosmotic pressure</span><span>, w</span><span>hile </span><span>antibiotics</span><span> is expensive and have </span><span>a bad</span><span> effect</span><span> on </span><span>environmental microbiology</span><span> because of drug resistance. However, the source to </span><span>hyperosmotic pressure</span><span> is </span><span>environmentally</span><span> friendly and won’t generate the risk of drug resistance. If our black hole system could be fully developed, it will </span><span>reduce the access barriers to microbiology research especially for the scientists from poor countries.</span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
 					    &nbsp;
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;">The source</span><span style="font-family: Times New Roman;">s</span><span style="font-family: Times New Roman;"> (such as NaCl and sucrose) to create hyperosmotic pressure</span><span style="font-family: Times New Roman;"> are cheap, </span><span style="font-family: Times New Roman;">accessible</span><span style="font-family: Times New Roman;"> and </span><span style="font-family: Times New Roman;">environmentally friendly</span><span style="font-family: Times New Roman;">, </span><span style="font-family: Times New Roman;">while antibiotics is expensive and have a bad effect on environmental microbiology because of drug resistance.</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">If our black hole system could be fully developed, barriers to microbiology research </span><span style="font-family: Times New Roman;">will be removed </span><span style="font-family: Times New Roman;">especially for the scientists from poor countries.</span>
+					    <span>The source</span><span>s</span><span> (such as NaCl and sucrose) to create hyperosmotic pressure</span><span> are cheap, </span><span>accessible</span><span> and </span><span>environmentally friendly</span><span>, </span><span>while antibiotics is expensive and have a bad effect on environmental microbiology because of drug resistance.</span><span> </span><span>If our black hole system could be fully developed, barriers to microbiology research </span><span>will be removed </span><span>especially for the scientists from poor countries.</span>
 					</p>
 				</div>
@@ Line 1,172: / Line 1,250: @@
 					    <span style="font-size: 18px;"></span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;">XMU-China 2013 has tried to </span><span style="font-family: Times New Roman;">construct </span><span style="font-family: Times New Roman;">o</span><span style="font-family: Times New Roman;">scillation </span><span style="font-family: Times New Roman;">system by standard biobricks. </span><span style="font-family: Times New Roman;">The synchronized oscillation syst</span><span style="font-family: Times New Roman;">em used in that study (Figure 1</span><span style="font-family: Times New Roman;">A</span><span style="font-family: Times New Roman;">) is based on the quorum sensing machineries in </span><span style="font-family: Times New Roman; font-style: italic;">Vibrio fischeri</span><span style="font-family: Times New Roman;"> and </span><span style="font-family: Times New Roman; font-style: italic;">Bacillus thurigensis</span><span style="font-family: Times New Roman;">. Three identical luxI promoters are in charge of </span><span style="font-family: Times New Roman; font-style: italic;">luxI</span><span style="font-family: Times New Roman;"> (from </span><span style="font-family: Times New Roman; font-style: italic;">V. fischeri</span><span style="font-family: Times New Roman;">), </span><span style="font-family: Times New Roman; font-style: italic;">aiiA</span><span style="font-family: Times New Roman;"> (from </span><span style="font-family: Times New Roman; font-style: italic;">B.thurigensis</span><span style="font-family: Times New Roman;">) and </span><span style="font-family: Times New Roman; font-style: italic;">gfp</span><span style="font-family: Times New Roman;"> genes separately. The LuxI synthase generates an acyl-homoserine-lactone (AHL), which can spread across the cell membrane and mediate intercellular coupling. AHL then binds </span><span style="font-family: Times New Roman;">to LuxR produced intracellularly</span><span style="font-family: Times New Roman;">, and the LuxR-AHL complex would activate the luxI promoter. AiiA catalyzes the degradation of AHL as the negative feedback in the circuit. Therefore, both the activator AHL and the repressor AiiA of the network are activated by the luxI</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">promoter simultaneously.</span>
+					    <span>XMU-China 2013 has tried to </span><span>construct </span><span>o</span><span>scillation </span><span>system by standard biobricks. </span><span>The synchronized oscillation syst</span><span>em used in that study (Figure 1</span><span>A</span><span>) is based on the quorum sensing machineries in </span><span style="font-family: ; font-style: italic;">Vibrio fischeri</span><span> and </span><span style="font-family: ; font-style: italic;">Bacillus thurigensis</span><span>. Three identical luxI promoters are in charge of </span><span style="font-family: ; font-style: italic;">luxI</span><span> (from </span><span style="font-family: ; font-style: italic;">V. fischeri</span><span>), </span><span style="font-family: ; font-style: italic;">aiiA</span><span> (from </span><span style="font-family: ; font-style: italic;">B.thurigensis</span><span>) and </span><span style="font-family: ; font-style: italic;">gfp</span><span> genes separately. The LuxI synthase generates an acyl-homoserine-lactone (AHL), which can spread across the cell membrane and mediate intercellular coupling. AHL then binds </span><span>to LuxR produced intracellularly</span><span>, and the LuxR-AHL complex would activate the luxI promoter. AiiA catalyzes the degradation of AHL as the negative feedback in the circuit. Therefore, both the activator AHL and the repressor AiiA of the network are activated by the luxI</span><span> </span><span>promoter simultaneously.</span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;"> </span>
+					    <span> </span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
 					    &nbsp;
 					</p>
@@ Line 1,185: / Line 1,263: @@
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 					            <td style="padding: 0px 7px; border: 1px rgb(0, 0, 0); border-image: none; width: 302px; vertical-align: top;">
-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <span style="font-family: Times New Roman; font-weight: 700;">A</span>
+					                    <span style="font-family: ; font-weight: 700;">A</span>
 					                </p>
-					                <p style="font-family: Times New Roman;">
+					                <p>
 					                    <img width="279" height="244" src="https://static.igem.org/mediawiki/2014/8/8a/Xmu_project_Oscillation01.png"/>
 					                </p>
@@ Line 1,200: / Line 1,278: @@
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 					            <td style="border-width: medium 1px 1px; border-color: currentColor rgb(0, 0, 0) rgb(0, 0, 0); padding: 0px 7px; width: 642px; vertical-align: top;" colspan="2">
-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <span style="font-family: Times New Roman;">Figure 1</span><span style="font-family: Times New Roman;"> A</span><span style="font-family: Times New Roman;"> </span>
+					                    <span>Figure 1</span><span> A</span><span> </span>
 					                </p>
-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <span style="font-family: Times New Roman;">B </span><span style="font-family: Times New Roman;">Two</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">oscillation cycles were observed within 500 minutes.</span>
+					                    <span>B </span><span>Two</span><span> </span><span>oscillation cycles were observed within 500 minutes.</span>
 					                </p>
 					            </td>
@@ Line 1,214: / Line 1,292: @@
 					    </tbody>
 					</table>
-					<p style="font-family: Times New Roman;">
+					<p>
 					    &nbsp;
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;">Based on above principle, one published paper has already realized </span><span style="font-family: Times New Roman;">synchronized oscillations</span><span style="font-family: Times New Roman;"> under </span><span style="font-family: Times New Roman;">microfluidic</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">device. However</span><span style="font-family: Times New Roman;">,</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">XMU-China 201</span><span style="font-family: Times New Roman;">3 can’</span><span style="font-family: Times New Roman;">t get </span><span style="font-family: Times New Roman;">synchronized </span><span style="font-family: Times New Roman;">oscillation</span><span style="font-family: Times New Roman;"> on microfluidics</span><span style="font-family: Times New Roman;">, and that will be </span><span style="font-family: Times New Roman;">discuss</span><span style="font-family: Times New Roman;">ed later. </span><span style="font-family: Times New Roman;">Through calculating </span><span style="font-family: Times New Roman;">fluorescence </span><span style="font-family: Times New Roman;">on 96-microwell plate e</span><span style="font-family: Times New Roman;">very 15 minutes, they got two oscillation cycles within 500 minutes (</span><span style="font-family: Times New Roman;">Figure 1B</span><span style="font-family: Times New Roman;">).</span>
+					    <span>Based on above principle, one published paper has already realized </span><span>synchronized oscillations</span><span> under </span><span>microfluidic</span><span> </span><span>device. However</span><span>,</span><span> </span><span>XMU-China 201</span><span>3 can’</span><span>t get </span><span>synchronized </span><span>oscillation</span><span> on microfluidics</span><span>, and that will be </span><span>discuss</span><span>ed later. </span><span>Through calculating </span><span>fluorescence </span><span>on 96-microwell plate e</span><span>very 15 minutes, they got two oscillation cycles within 500 minutes (</span><span>Figure 1B</span><span>).</span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
 					    &nbsp;
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;">Based on that, we con</span><span style="font-family: Times New Roman;">struct our circuit by replacing </span><span style="font-family: Times New Roman; font-style: italic;">GFP</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">with</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman; font-style: italic;">CheZ</span><span style="font-family: Times New Roman; font-style: italic;"> </span><span style="font-family: Times New Roman;">(Figure 2)</span><span style="font-family: Times New Roman;">.</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">As the expression strength of </span><span style="font-family: Times New Roman; font-style: italic;">CheZ</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">is </span><span style="font-family: Times New Roman;">oscillatory</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">fluctuating</span><span style="font-family: Times New Roman;">, </span><span style="font-family: Times New Roman;">the motile ability will change </span><span style="font-family: Times New Roman;">periodically</span><span style="font-family: Times New Roman;">.</span><span style="font-family: Times New Roman;"> Bacteria will have the strongest motile ability at wave crest while even be non-motile at wave tr</span><span style="font-family: Times New Roman;">ough. Thus, the periodical </span><span style="font-family: Times New Roman;">change of motile ability leads to bacteria density </span><span style="font-family: Times New Roman;">distributing </span><span style="font-family: Times New Roman;">unevenly. When the bacteria </span><span style="font-family: Times New Roman;">are</span><span style="font-family: Times New Roman;"> at non-motile period, </span><span style="font-family: Times New Roman;">they</span><span style="font-family: Times New Roman;"> will aggregate together leading to the formation of </span><span style="font-family: Times New Roman;">growth</span><span style="font-family: Times New Roman;">-ring</span><span style="font-family: Times New Roman;">-like</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">patterns which could be distinguished by naked eyes.</span>
+					    <span>Based on that, we con</span><span>struct our circuit by replacing </span><span style="font-family: ; font-style: italic;">GFP</span><span> </span><span>with</span><span> </span><span style="font-family: ; font-style: italic;">CheZ</span><span style="font-family: ; font-style: italic;"> </span><span>(Figure 2)</span><span>.</span><span> </span><span>As the expression strength of </span><span style="font-family: ; font-style: italic;">CheZ</span><span> </span><span>is </span><span>oscillatory</span><span> </span><span>fluctuating</span><span>, </span><span>the motile ability will change </span><span>periodically</span><span>.</span><span> Bacteria will have the strongest motile ability at wave crest while even be non-motile at wave tr</span><span>ough. Thus, the periodical </span><span>change of motile ability leads to bacteria density </span><span>distributing </span><span>unevenly. When the bacteria </span><span>are</span><span> at non-motile period, </span><span>they</span><span> will aggregate together leading to the formation of </span><span>growth</span><span>-ring</span><span>-like</span><span> </span><span>patterns which could be distinguished by naked eyes.</span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
 					    &nbsp;
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;">Many trees in </span><span style="font-family: Times New Roman;">temperate</span><span style="font-family: Times New Roman;"> zones make one growth ring each year, </span><span style="font-family: Times New Roman;">with the newest adjacent to the bark</span><span style="font-family: Times New Roman;">. We can tell </span><span style="font-family: Times New Roman;">a tree’s </span><span style="font-family: Times New Roman;">age b</span><span style="font-family: Times New Roman;">y counting the number of growth </span><span style="font-family: Times New Roman;">ring</span><span style="font-family: Times New Roman;">s</span><span style="font-family: Times New Roman;">. A</span><span style="font-family: Times New Roman;">nalogously</span><span style="font-family: Times New Roman;">, bacteria rings could also be formed by gene </span><span style="font-family: Times New Roman;">oscillator</span><span style="font-family: Times New Roman;">. </span><span style="font-family: Times New Roman;">Multiply</span><span style="font-family: Times New Roman;"> the </span><span style="font-family: Times New Roman;">period</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">by</span><span style="font-family: Times New Roman;"> the </span><span style="font-family: Times New Roman;">quantity</span><span style="font-family: Times New Roman;"> of bacteria rings, we c</span><span style="font-family: Times New Roman;">an tell how much time has passed.</span>
+					    <span>Many trees in </span><span>temperate</span><span> zones make one growth ring each year, </span><span>with the newest adjacent to the bark</span><span>. We can tell </span><span>a tree’s </span><span>age b</span><span>y counting the number of growth </span><span>ring</span><span>s</span><span>. A</span><span>nalogously</span><span>, bacteria rings could also be formed by gene </span><span>oscillator</span><span>. </span><span>Multiply</span><span> the </span><span>period</span><span> </span><span>by</span><span> the </span><span>quantity</span><span> of bacteria rings, we c</span><span>an tell how much time has passed.</span>
 					</p>
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@@ Line 1,236: / Line 1,314: @@
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-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <img width="356" height="310" style="font-family: Times New Roman;" src="https://static.igem.org/mediawiki/2014/6/63/Xmu_project_Oscillation03.png"/>
+					                    <img width="356" height="310" src="https://static.igem.org/mediawiki/2014/6/63/Xmu_project_Oscillation03.png"/>
 					                </p>
 					            </td>
@@ Line 1,243: / Line 1,321: @@
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-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <span style="font-family: Times New Roman;">Figure</span><span style="font-family: Times New Roman;"> 2</span>
+					                    <span>Figure</span><span> 2</span>
 					                </p>
 					            </td>
@@ Line 1,253: / Line 1,331: @@
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 					</table>
-					<p style="font-family: Times New Roman;">
+					<p>
 					    &nbsp;
 					</p>
@@ Line 1,262: / Line 1,340: @@
 				<img id="Project_Psystem_title" class="Project_title" src="https://static.igem.org/mediawiki/2014/a/a9/Xmu_project_background.png"  />
 				<div id="Project_Psystem_wd">
-					<p style="text-align: justify; font-family: Times New Roman; styleName: Normal;">
+					<p style="text-align: justify; font-family: ; styleName: Normal;">
-					    <span style="font-family: Times New Roman;">In</span><span style="font-family: Times New Roman;"> the project </span><span style="font-family: Times New Roman;">of</span><span style="font-family: Times New Roman;"> iGEM</span><span style="font-family: Times New Roman;">13 XMU-China, they</span><span style="font-family: Times New Roman;"> can’t get expected oscillation. H</span><span style="font-family: Times New Roman;">owever</span><span style="font-family: Times New Roman;">,</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">this year</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">iGEM14 XMU-China</span><span style="font-family: Times New Roman;"> further </span><span style="font-family: Times New Roman;">investigate the reason of abnormal oscillation. We further review</span><span style="font-family: Times New Roman;"> SDS-PAGE analysis to confirm </span><span style="font-family: Times New Roman;">the circuit </span><span style="font-family: Times New Roman;">at</span><span style="font-family: Times New Roman;"> protein leve</span><span style="font-family: Times New Roman;">l. The SDS-PAGE data is shown in </span><span style="font-family: Times New Roman; font-weight: 700;">Figure 1</span><span style="font-family: Times New Roman;">.</span><span style="font-family: Times New Roman;"> Based on that, we make a reasonable assumption that the unexpected behavior of the LuxR Promoter lead</span><span style="font-family: Times New Roman;">s</span><span style="font-family: Times New Roman;"> to the misfolding proteins </span><span style="font-family: Times New Roman; font-size: 14px; styleName: Default Paragraph Font;"></span><span style="font-family: Times New Roman;">hence the</span><span style="font-family: Times New Roman;"> abnormal oscillation.</span>
+					    <span>In</span><span> the project </span><span>of</span><span> iGEM</span><span>13 XMU-China, they</span><span> can’t get expected oscillation. H</span><span>owever</span><span>,</span><span> </span><span>this year</span><span> </span><span>iGEM14 XMU-China</span><span> further </span><span>investigate the reason of abnormal oscillation. We further review</span><span> SDS-PAGE analysis to confirm </span><span>the circuit </span><span>at</span><span> protein leve</span><span>l. The SDS-PAGE data is shown in </span><span style="font-family: ; font-weight: 700;">Figure 1</span><span>.</span><span> Based on that, we make a reasonable assumption that the unexpected behavior of the LuxR Promoter lead</span><span>s</span><span> to the misfolding proteins </span><span style="font-family: ; font-size: 14px; styleName: Default Paragraph Font;"></span><span>hence the</span><span> abnormal oscillation.</span>
 					</p>
-					<p style="text-align: justify; font-family: Times New Roman; styleName: Normal;">
+					<p style="text-align: justify; font-family: ; styleName: Normal;">
 					    &nbsp;
 					</p>
@@ Line 1,272: / Line 1,350: @@
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-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <img width="600" style="font-family: Times New Roman;" src="https://static.igem.org/mediawiki/2014/2/22/Xmu_project_p_system01.png"/>
+					                    <img width="600" src="https://static.igem.org/mediawiki/2014/2/22/Xmu_project_p_system01.png"/>
 					                </p>
 					            </td>
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-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <span style="font-family: Times New Roman; font-weight: 700;">Figure 1</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">SDS-PAGE analysis of E.</span><span style="font-family: Times New Roman;">coli K strain (</span><span style="font-family: Times New Roman; font-style: italic;">DH5α</span><span style="font-family: Times New Roman;">). </span><span style="font-family: Times New Roman; font-weight: 700;">(a)</span><span style="font-family: Times New Roman;"> Lane 1-2: supernatant and pellet of original </span><span style="font-family: Times New Roman; font-style: italic;">DH5α</span><span style="font-family: Times New Roman;">; Lane 3-4: supernatant and pellet of strain with single plasmid A1 (BBa_K1036003); Lane 5-6: supernatant and pellet of strain with both plasmids A1</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">(BBa_K1036003) and B</span><span style="font-family: Times New Roman;"> (BBa_K1036000)</span><span style="font-family: Times New Roman;">. The red arrows indicate the </span><span style="font-family: Times New Roman; font-weight: 700;">misfolding</span><span style="font-family: Times New Roman;"> GFP-LVA protein (27.6 kDa) in the precipitation. </span><span style="font-family: Times New Roman; font-weight: 700;">(b)</span><span style="font-family: Times New Roman;"> Lane 1-2: supernatant and pellet of original </span><span style="font-family: Times New Roman; font-style: italic;">BL21</span><span style="font-family: Times New Roman;">; Lane 3-4: supernatant and pellet of strain with single plasmid A1</span><span style="font-family: Times New Roman;"> (BBa_K1036003)</span><span style="font-family: Times New Roman;">; Lane 5-6: supernatant and pellet of strain with both plasmids A1</span><span style="font-family: Times New Roman;"> (BBa_K1036003) </span><span style="font-family: Times New Roman;">and B</span><span style="font-family: Times New Roman;"> (BBa_K1036000)</span><span style="font-family: Times New Roman;">. The blue arrows indicate LuxR (27.5 kDa), GFP-LVA (27.6 kDa) and AiiA-LVA (28.7 kDa) in the supernatant. The orange arrows indicate LuxI-LVA (22.4 kDa) in the supernatant. (The marker of b was not in right position, however, the proteins were confirmed by MALDI-TOF-TOF .)</span>
+					                    <span style="font-family: ; font-weight: 700;">Figure 1</span><span> </span><span>SDS-PAGE analysis of E.</span><span>coli K strain (</span><span style="font-family: ; font-style: italic;">DH5α</span><span>). </span><span style="font-family: ; font-weight: 700;">(a)</span><span> Lane 1-2: supernatant and pellet of original </span><span style="font-family: ; font-style: italic;">DH5α</span><span>; Lane 3-4: supernatant and pellet of strain with single plasmid A1 (BBa_K1036003); Lane 5-6: supernatant and pellet of strain with both plasmids A1</span><span> </span><span>(BBa_K1036003) and B</span><span> (BBa_K1036000)</span><span>. The red arrows indicate the </span><span style="font-family: ; font-weight: 700;">misfolding</span><span> GFP-LVA protein (27.6 kDa) in the precipitation. </span><span style="font-family: ; font-weight: 700;">(b)</span><span> Lane 1-2: supernatant and pellet of original </span><span style="font-family: ; font-style: italic;">BL21</span><span>; Lane 3-4: supernatant and pellet of strain with single plasmid A1</span><span> (BBa_K1036003)</span><span>; Lane 5-6: supernatant and pellet of strain with both plasmids A1</span><span> (BBa_K1036003) </span><span>and B</span><span> (BBa_K1036000)</span><span>. The blue arrows indicate LuxR (27.5 kDa), GFP-LVA (27.6 kDa) and AiiA-LVA (28.7 kDa) in the supernatant. The orange arrows indicate LuxI-LVA (22.4 kDa) in the supernatant. (The marker of b was not in right position, however, the proteins were confirmed by MALDI-TOF-TOF .)</span>
 					                </p>
 					            </td>
@@ Line 1,289: / Line 1,367: @@
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 					</table>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;">As the SDS-PAGE show</span><span style="font-family: Times New Roman;">s</span><span style="font-family: Times New Roman;">, a large amount of </span><span style="font-family: Times New Roman;">GFP-LVA and LuxI-LVA </span><span style="font-family: Times New Roman;">appear</span><span style="font-family: Times New Roman;"> in pellet where misfolding proteins often exist. Both protein</span><span style="font-family: Times New Roman;">s</span><span style="font-family: Times New Roman;"> directl</span><span style="font-family: Times New Roman;">y </span><span style="font-family: Times New Roman;">a</span><span style="font-family: Times New Roman;">ffect the oscillation result. </span><span style="font-family: Times New Roman;">And it is critical to find out the reason for misfolding proteins</span><span style="font-family: Times New Roman;">. iGEM14 XMU-China make the following assumption:</span>
+					    <span>As the SDS-PAGE show</span><span>s</span><span>, a large amount of </span><span>GFP-LVA and LuxI-LVA </span><span>appear</span><span> in pellet where misfolding proteins often exist. Both protein</span><span>s</span><span> directl</span><span>y </span><span>a</span><span>ffect the oscillation result. </span><span>And it is critical to find out the reason for misfolding proteins</span><span>. iGEM14 XMU-China make the following assumption:</span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;"> &nbsp; &nbsp;</span>
+					    <span> &nbsp; &nbsp;</span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman;">The 2012 published paper reveals an unexpected behavior of </span><span style="font-family: Times New Roman;">L</span><span style="font-family: Times New Roman;">ux pR</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">(BBa_R0062).</span><span style="font-family: Times New Roman;"> In the absence of autoinducer 3OC6 (AHL), LuxR binds to Plux</span><span style="font-family: Times New Roman;"> (Lux pR)</span><span style="font-family: Times New Roman;"> and activates backwards transcription (</span><span style="font-family: Times New Roman; font-weight: 700;">Figure 2</span><span style="font-family: Times New Roman;">).</span>
+					    <span>The 2012 published paper reveals an unexpected behavior of </span><span>L</span><span>ux pR</span><span> </span><span>(BBa_R0062).</span><span> In the absence of autoinducer 3OC6 (AHL), LuxR binds to Plux</span><span> (Lux pR)</span><span> and activates backwards transcription (</span><span style="font-family: ; font-weight: 700;">Figure 2</span><span>).</span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
 					    &nbsp;
 					</p>
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-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <img width="540" height="409" style="font-family: Times New Roman;" src="https://static.igem.org/mediawiki/2014/c/c5/Xmu_project_p_system02.png"/>
+					                    <img width="540" height="409" src="https://static.igem.org/mediawiki/2014/c/c5/Xmu_project_p_system02.png"/>
 					                </p>
 					            </td>
@@ Line 1,312: / Line 1,390: @@
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 					            <td style="border-width: medium 1px 1px; border-color: currentColor rgb(0, 0, 0) rgb(0, 0, 0); padding: 0px 7px; width: 567px; vertical-align: top;">
-					                <p style="font-family: Times New Roman;">
+					                <p>
-					                    <span style="font-family: Times New Roman; font-weight: 700;">Figure</span><span style="font-family: Times New Roman; font-weight: 700;"> 2</span><span style="font-family: Times New Roman;"> Relative RFP fluorescence for a control construct designed to measure backwards transcription from </span><span style="font-family: Times New Roman;">Lux pR</span><span style="font-family: Times New Roman;">. Addition of LuxR and 3OC6</span><span style="font-family: Times New Roman;">90 (AHL)</span><span style="font-family: Times New Roman;"> as indicated. Error bars in all panels are one standard deviation.</span>
+					                    <span style="font-family: ; font-weight: 700;">Figure</span><span style="font-family: ; font-weight: 700;"> 2</span><span> Relative RFP fluorescence for a control construct designed to measure backwards transcription from </span><span>Lux pR</span><span>. Addition of LuxR and 3OC6</span><span>90 (AHL)</span><span> as indicated. Error bars in all panels are one standard deviation.</span>
 					                </p>
 					            </td>
@@ Line 1,322: / Line 1,400: @@
 					    </tbody>
 					</table>
-					<p style="font-family: Times New Roman;">
+					<p>
 					    &nbsp;
 					</p>
-					<p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
+					<p style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">
-					    <span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">The imperfect simplification</span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;"> of setting </span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">lux pL</span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;"> and </span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">L</span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">ux pR</span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;"> in the same direction</span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">:</span>
+					    <span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">The imperfect simplification</span><span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;"> of setting </span><span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">lux pL</span><span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;"> and </span><span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">L</span><span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">ux pR</span><span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;"> in the same direction</span><span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">:</span>
 					</p>
-					<p style="text-align: justify; font-family: Times New Roman; font-weight: 700; styleName: Normal;">
+					<p style="text-align: justify; font-family: ; font-weight: 700; styleName: Normal;">
-					    <span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">From the original design by</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">Jeff</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">H</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">asty</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">, we can see that Lux pR and Lux pL are </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">set</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> in opposite directions</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> (</span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">Figure 3</span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">A</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">)</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">.</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">In the absence of AHL</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">, LuxR could activate backward transcription of Lux pR</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: Times New Roman; font-size: 14px; styleName: Default Paragraph Font;"></span><span style="font-family: Times New Roman;">leading to more expression of LuxR which is</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> critical to meet the oscillation conditions. However, present literature don’t consider </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">the backwards transcription which </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">have effect</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> on </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">q</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">uorum sensing oscillation.</span>
+					    <span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">From the original design by</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">Jeff</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">H</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">asty</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">, we can see that Lux pR and Lux pL are </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">set</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> in opposite directions</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> (</span><span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">Figure 3</span><span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">A</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">)</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">.</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">In the absence of AHL</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">, LuxR could activate backward transcription of Lux pR</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: ; font-size: 14px; styleName: Default Paragraph Font;"></span><span>leading to more expression of LuxR which is</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> critical to meet the oscillation conditions. However, present literature don’t consider </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">the backwards transcription which </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">have effect</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> on </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">q</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">uorum sensing oscillation.</span>
 					</p>
-					<p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
+					<p style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">
 					    &nbsp;
 					</p>
@@ Line 1,338: / Line 1,416: @@
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-					                <p style="text-align: justify; text-indent: -24px; font-family: Times New Roman; font-weight: 700; margin-left: 24px; styleName: Normal;">
+					                <p style="text-align: justify; text-indent: -24px; font-family: ; font-weight: 700; margin-left: 24px; styleName: Normal;">
-					                    <span style="font-weight: 700;">A.</span><span style="font-weight: 700;">	</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">Original </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">D</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">esign</span>
+					                    <span style="font-weight: 700;">A.</span><span style="font-weight: 700;">	</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">Original </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">D</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">esign</span>
 					                </p>
-					                <p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
+					                <p style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">
-					                    <img width="535" height="283" style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;" src="https://static.igem.org/mediawiki/2014/3/30/Xmu_project_p_system03.png"/>
+					                    <img width="535" height="283" style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;" src="https://static.igem.org/mediawiki/2014/3/30/Xmu_project_p_system03.png"/>
 					                </p>
 					            </td>
@@ Line 1,348: / Line 1,426: @@
 					        <tr>
 					            <td style="border-width: medium 1px 1px; border-color: currentColor rgb(0, 0, 0) rgb(0, 0, 0); padding: 0px 7px; width: 553px; vertical-align: top;">
-					                <p style="text-align: justify; text-indent: -24px; font-family: Times New Roman; font-weight: 700; margin-left: 24px; styleName: Normal;">
+					                <p style="text-align: justify; text-indent: -24px; font-family: ; font-weight: 700; margin-left: 24px; styleName: Normal;">
-					                    <span style="font-weight: 700;">B.</span><span style="font-weight: 700;">	</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">iGEM13 XMU-China Design</span>
+					                    <span style="font-weight: 700;">B.</span><span style="font-weight: 700;">	</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">iGEM13 XMU-China Design</span>
 					                </p>
-					                <p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
+					                <p style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">
-					                    <img width="543" height="250" style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;" src="https://static.igem.org/mediawiki/2014/3/35/Xmu_project_p_system04.png"/>
+					                    <img width="543" height="250" style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;" src="https://static.igem.org/mediawiki/2014/3/35/Xmu_project_p_system04.png"/>
 					                </p>
 					            </td>
@@ Line 1,358: / Line 1,436: @@
 					        <tr>
 					            <td style="border-width: medium 1px 1px; border-color: currentColor rgb(0, 0, 0) rgb(0, 0, 0); padding: 0px 7px; width: 553px; vertical-align: top;">
-					                <p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
+					                <p style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">
-					                    <span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">Figure 3</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">A.</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">Top row is the original design by</span><span style="font-size: 14px;"> </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">Jeff Hasty</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">.</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">B.</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">Bottom row is the simplif</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">ied</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> design which sets lux pL and lux pR in the same direction.</span>
+					                    <span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">Figure 3</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">A.</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">Top row is the original design by</span><span style="font-size: 14px;"> </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">Jeff Hasty</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">.</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">B.</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">Bottom row is the simplif</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">ied</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> design which sets lux pL and lux pR in the same direction.</span>
 					                </p>
 					            </td>
@@ Line 1,368: / Line 1,446: @@
 					    </tbody>
 					</table>
-					<p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
+					<p style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">
-					    <span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">In the simplif</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">ied</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> design (</span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">Figure 3B</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">), when LuxR activates the backward transcription, RNA polymerase will be blocked by the terminators B0015. So that thi</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">s simplif</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">ication</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">doesn’t follow the original design. Actually, the</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> reverse </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">terminated </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">efficiency </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">of B0015 is 0.295</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">(CC) which may lead to leakage transcription. </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">However, the correct sequence of GFP-LAA can’t be transcribed during the backwards transcription, even if the minus-strand of GFP-LAA could be transcribed, the sequence of the RNA is not the right codons of GFP-LAA. hence incorrect amino acid sequences may be translated, resulting in misfolding GFP expression which is just as the SDS-PAGE shows (</span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">Figure 1</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">). </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">Misfolding GFP may also be produced by </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">bi-directional</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> transcription</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">.</span>
+					    <span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">In the simplif</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">ied</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> design (</span><span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">Figure 3B</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">), when LuxR activates the backward transcription, RNA polymerase will be blocked by the terminators B0015. So that thi</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">s simplif</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">ication</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">doesn’t follow the original design. Actually, the</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> reverse </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">terminated </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">efficiency </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">of B0015 is 0.295</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">(CC) which may lead to leakage transcription. </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">However, the correct sequence of GFP-LAA can’t be transcribed during the backwards transcription, even if the minus-strand of GFP-LAA could be transcribed, the sequence of the RNA is not the right codons of GFP-LAA. hence incorrect amino acid sequences may be translated, resulting in misfolding GFP expression which is just as the SDS-PAGE shows (</span><span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">Figure 1</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">). </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">Misfolding GFP may also be produced by </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">bi-directional</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> transcription</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">.</span>
 					</p>
-					<p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
+					<p style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">
 					    &nbsp;
 					</p>
-					<p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
+					<p style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">
-					    <span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">Because of the </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">imperfect simplified</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> design doesn’t follow the original function completely, the abnormal oscillation is justifiable. The misfolding protein is a evidence to support our assumption.</span>
+					    <span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">Because of the </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">imperfect simplified</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> design doesn’t follow the original function completely, the abnormal oscillation is justifiable. The misfolding protein is a evidence to support our assumption.</span>
 					</p>
-					<p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
+					<p style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">
 					    &nbsp;
 					</p>
-					<p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
+					<p style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">
-					    <span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">iGEM14 XMU-China involved sequence comparison to investigate the difference between the </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">original and </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">the registry parts.</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> We find that</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> the original Lux pR has 20bp overlapping sequence with origin</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">al Lux p</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">R</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">. </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">There is </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">a r</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">estriction e</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">nzyme cutting site (EcoR I)</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> at the 56bp of original Lux pR</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> (</span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">Figure 4</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">)</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">.</span>
+					    <span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">iGEM14 XMU-China involved sequence comparison to investigate the difference between the </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">original and </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">the registry parts.</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> We find that</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> the original Lux pR has 20bp overlapping sequence with origin</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">al Lux p</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">R</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">. </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">There is </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">a r</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">estriction e</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">nzyme cutting site (EcoR I)</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> at the 56bp of original Lux pR</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> (</span><span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">Figure 4</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">)</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">.</span>
 					</p>
 					<table  width="600px" style="position:relative;margin-left:-300px;left:50%">
@@ Line 1,387: / Line 1,465: @@
 					        <tr class="firstRow">
 					            <td style="padding: 0px 7px; border: 1px rgb(0, 0, 0); width: 245px; vertical-align: top;">
-					                <p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
+					                <p style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">
-					                    <img width="225" height="255" style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;" src="https://static.igem.org/mediawiki/2014/0/00/Xmu_project_p_system05.png"/>
+					                    <img width="225" height="255" style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;" src="https://static.igem.org/mediawiki/2014/0/00/Xmu_project_p_system05.png"/>
 					                </p>
 					            </td>
@@ Line 1,394: / Line 1,472: @@
 					        <tr>
 					            <td style="border-width: medium 1px 1px; border-color: currentColor rgb(0, 0, 0) rgb(0, 0, 0); padding: 0px 7px; width: 245px; vertical-align: top;">
-					                <p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
+					                <p style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">
-					                    <span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">Figure 4</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> Schematic of original P system.</span>
+					                    <span style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">Figure 4</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> Schematic of original P system.</span>
 					                </p>
 					            </td>
@@ Line 1,404: / Line 1,482: @@
 					    </tbody>
 					</table>
-					<p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
+					<p style="font-family: ; font-weight: 700; styleName: Default Paragraph Font;">
-					    <span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> </span>
+					    <span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> </span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    <span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">Parts registry truncate the original Lux pR at 56bp to get the </span><span style="font-family: Times New Roman;">55bp Lux pR</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">(BBa_R0062). </span><span style="font-family: Times New Roman;">On the contrary, Lux pL</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">(BBa_R0063) is longer the original Lux pL, and at the end of BBa_R0063 is </span><span style="font-family: Times New Roman;">initial part of </span><span style="font-family: Times New Roman;">41bp </span><span style="font-family: Times New Roman;">LuxR (BBa_C0062). </span><span style="font-family: Times New Roman;">Thus new problems</span><span style="font-family: Times New Roman;"> arise</span><span style="font-family: Times New Roman;">—Is the modification of original P system reasonable? Does the m</span><span style="font-family: Times New Roman;">odification result in the unexpected backward transcription?</span>
+					    <span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">Parts registry truncate the original Lux pR at 56bp to get the </span><span>55bp Lux pR</span><span> </span><span>(BBa_R0062). </span><span>On the contrary, Lux pL</span><span> </span><span>(BBa_R0063) is longer the original Lux pL, and at the end of BBa_R0063 is </span><span>initial part of </span><span>41bp </span><span>LuxR (BBa_C0062). </span><span>Thus new problems</span><span> arise</span><span>—Is the modification of original P system reasonable? Does the m</span><span>odification result in the unexpected backward transcription?</span>
 					</p>
-					<p style="font-family: Times New Roman;">
+					<p>
-					    &nbsp;
-					</p>
-					<p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
-					    <span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">Quorum sensing system is so widely used in the synthetic biology, we think it’s remarkable to make it clear. We highlight the abnormal phenomenon of QS oscillation which may</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> be caused by imperfect simplification</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> for the very first time. </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">We hope</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> that more </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">effort</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> would be </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">made</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> to figure out the interaction between </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">QS oscillation parts.</span>
-					</p>
-					<p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
-					    &nbsp;
-					</p>
-				</div>
-			</div>
-		</div>
-	</body>
-</html>
-tyle="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">ied</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> design (</span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">Figure 3B</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">), when LuxR activates the backward transcription, RNA polymerase will be blocked by the terminators B0015. So that thi</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">s simplif</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">ication</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">doesn’t follow the original design. Actually, the</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> reverse </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">terminated </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">efficiency </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">of B0015 is 0.295</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">(CC) which may lead to leakage transcription. </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">However, the correct sequence of GFP-LAA can’t be transcribed during the backwards transcription, even if the minus-strand of GFP-LAA could be transcribed, the sequence of the RNA is not the right codons of GFP-LAA. hence incorrect amino acid sequences may be translated, resulting in misfolding GFP expression which is just as the SDS-PAGE shows (</span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">Figure 1</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">). </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">Misfolding GFP may also be produced by </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">bi-directional</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> transcription</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">.</span>
-					</p>
-					<p style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">
-					    &nbsp;
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-					    <span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">Because of the </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">imperfect simplified</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> design doesn’t follow the original function completely, the abnormal oscillation is justifiable. The misfolding protein is a evidence to support our assumption.</span>
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-					    <span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">iGEM14 XMU-China involved sequence comparison to investigate the difference between the </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">original and </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">the registry parts.</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> We find that</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> the original Lux pR has 20bp overlapping sequence with origin</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">al Lux p</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">R</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">. </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">There is </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">a r</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">estriction e</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">nzyme cutting site (EcoR I)</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> at the 56bp of original Lux pR</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> (</span><span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">Figure 4</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">)</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">.</span>
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-					                    <img width="225" height="255" style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;" src="https://static.igem.org/mediawiki/2014/0/00/Xmu_project_p_system05.png"/>
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-					                    <span style="font-family: Times New Roman; font-weight: 700; styleName: Default Paragraph Font;">Figure 4</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> Schematic of original P system.</span>
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-					    <span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">Parts registry truncate the original Lux pR at 56bp to get the </span><span style="font-family: Times New Roman;">55bp Lux pR</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">(BBa_R0062). </span><span style="font-family: Times New Roman;">On the contrary, Lux pL</span><span style="font-family: Times New Roman;"> </span><span style="font-family: Times New Roman;">(BBa_R0063) is longer the original Lux pL, and at the end of BBa_R0063 is </span><span style="font-family: Times New Roman;">initial part of </span><span style="font-family: Times New Roman;">41bp </span><span style="font-family: Times New Roman;">LuxR (BBa_C0062). </span><span style="font-family: Times New Roman;">Thus new problems</span><span style="font-family: Times New Roman;"> arise</span><span style="font-family: Times New Roman;">—Is the modification of original P system reasonable? Does the m</span><span style="font-family: Times New Roman;">odification result in the unexpected backward transcription?</span>
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-					    <span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">Quorum sensing system is so widely used in the synthetic biology, we think it’s remarkable to make it clear. We highlight the abnormal phenomenon of QS oscillation which may</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> be caused by imperfect simplification</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> for the very first time. </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">We hope</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> that more </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">effort</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> would be </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">made</span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;"> to figure out the interaction between </span><span style="font-family: Times New Roman; font-weight: 400; styleName: Default Paragraph Font;">QS oscillation parts.</span>
+					    <span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">Quorum sensing system is so widely used in the synthetic biology, we think it’s remarkable to make it clear. We highlight the abnormal phenomenon of QS oscillation which may</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> be caused by imperfect simplification</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> for the very first time. </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">We hope</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> that more </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">effort</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> would be </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">made</span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;"> to figure out the interaction between </span><span style="font-family: ; font-weight: 400; styleName: Default Paragraph Font;">QS oscillation parts.</span>
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