Team:NCTU Formosa/biobricks

From 2014.igem.org

(Difference between revisions)
(→Brief Information)
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Please click on the name of the parts for detailed information that is hosted in the Registry website.
Please click on the name of the parts for detailed information that is hosted in the Registry website.
====PBAN-producted system====
====PBAN-producted system====
-
======BBa_K1017726======
+
======PBAN1======
*pcya and ho1 are the enzymes needed to convert heme into chromophore phycocyanobiline(PCB), the red light sensor. Since pcya and ho1 are not naturally produced in E.coli, we use P<sub>cons</sub> upstream in order to make E.coli continuously expressed them to synthesize PCB.
*pcya and ho1 are the enzymes needed to convert heme into chromophore phycocyanobiline(PCB), the red light sensor. Since pcya and ho1 are not naturally produced in E.coli, we use P<sub>cons</sub> upstream in order to make E.coli continuously expressed them to synthesize PCB.
-
<br>[[File:Nctu_pcya_ho1.jpg|450px]]
+
<br>[[File:NCTU_Formosa_2014_Biobrick1|450px]]
-
======BBa_K1017301======
+
======PBAN2======
*Cph8 is a chimeric light receptor. It is a fusion of the photoreceptor cph1 and the envZ histidine kinase. cph1 is only active when it binds the chromophore phycocyanobiline (PCB).
*Cph8 is a chimeric light receptor. It is a fusion of the photoreceptor cph1 and the envZ histidine kinase. cph1 is only active when it binds the chromophore phycocyanobiline (PCB).
-
<br>[[File:Nctu_cph8.jpg|150px]]
+
<br>[[File:NCTU_Formosa_2014_Biobrick2|150px]]
-
======BBa_K1017781======
+
======PBAN3======
*This is the key part to change Ompc promoter intro red promotor. The lacI will inhibit the function of P<sub>lac</sub>, thus, the original outcomes will be converted into our expected result.
*This is the key part to change Ompc promoter intro red promotor. The lacI will inhibit the function of P<sub>lac</sub>, thus, the original outcomes will be converted into our expected result.
-
<br>[[File:Nctu_lacI_plac.jpg|300px]]
+
<br>[[File:NCTU_Formosa_2014_Biobrick3|300px]]
-
======BBa_K1017101======
+
======PBAN4======
*Ompc promtor, which can sense red light with the presence of cph8. It will be turned on in the dark ,and be turned off in the bright. For the convenient use, we add lacI and lac promotor downstream the biobrick. This part, so called red promotor, can be activated under red light, and inactive in the dark.  
*Ompc promtor, which can sense red light with the presence of cph8. It will be turned on in the dark ,and be turned off in the bright. For the convenient use, we add lacI and lac promotor downstream the biobrick. This part, so called red promotor, can be activated under red light, and inactive in the dark.  
-
<br>[[File:Nctu_Pred.jpg|450px]]
+
<br>[[File:NCTU_Formosa_2014_Biobrick4|450px]]
-
====Temperature-regulated system====
+
======PBAN5======
-
======BBa_K1017602======
+
<br>[[File:NCTU_Formosa_2014_Biobrick5|200px]]
 +
======PBAN6======
*By using mGFP as a reporter gene, we can test whether the 37 °C RBS works.
*By using mGFP as a reporter gene, we can test whether the 37 °C RBS works.
-
<br>[[File:Nctu_37rbs_mGFP.jpg|200px]]
+
<br>[[File:NCTU_Formosa_2014_Biobrick6|200px]]
-
======BBa_K1017603======
+
======PBAN7======
*In our circuit, this biobrick is the part of P<sub>lux</sub>'s activation when the temperature reaches to 37<sup>o</sup>C.
*In our circuit, this biobrick is the part of P<sub>lux</sub>'s activation when the temperature reaches to 37<sup>o</sup>C.
<br>
<br>
-
[[File:Nctu_37rbs_luxr.jpg|200px]]
+
[[File:NCTU_Formosa_2014_Biobrick7|200px]]
-
====Small RNA-regulated system====
 
-
======BBa_K1017403======
+
======PBAN8======
*The sRNA is the complement of its rRBS. It can regulate the downstream of rRBS in RNA level by binding onto the rRBS when it is transcribed in order to interrupt ribosomes' work. In addition, adding Plux upstream makes the sequence be controlled by luxR/AHL complex.<br>
*The sRNA is the complement of its rRBS. It can regulate the downstream of rRBS in RNA level by binding onto the rRBS when it is transcribed in order to interrupt ribosomes' work. In addition, adding Plux upstream makes the sequence be controlled by luxR/AHL complex.<br>
-
[[File:Nctu_plux_srna1.jpg|250px]]
+
[[File:NCTU_Formosa_2014_Biobrick8|250px]]
-
======BBa_K1017404======
+
======PBAN9======
-
*The sRNA is the complement of its rRBS. It can regulate the downstream of rRBS in RNA level by binding onto the rRBS when it is transcribed in order to interrupt ribosomes' work.<br>[[File:Nctu_srna2.jpg|200px]]
+
*The sRNA is the complement of its rRBS. It can regulate the downstream of rRBS in RNA level by binding onto the rRBS when it is transcribed in order to interrupt ribosomes' work.
 +
<br>[[File:NCTU_Formosa_2014_Biobrick9|200px]]
-
======BBa_K1017202======
 
-
*The sRNA, base pair with target mRNA, including the Shine-Dalgarno sequence. Thus it prevent ribosome from binding to initiate the translation. The rRBS is designed for sRNA perfect binding, and this rRBS is the RBS which can be bound only for our artificial sRNA(BBa_K1017404).<br>
 
-
[[File:Nctu_rbs2.jpg|150px]]
 
-
 
-
======BBa_K1017811======
 
-
*The sequence let us show the efficiency of the sRNA-rRBS binding by expressing the fluorescence with the combination of a sequence providing the transcription of the sRNA.
 
-
[[File:Nctu_pcons_rbs2_mrfp.jpg|350px]]
 
-
 
-
======BBa_K1017401======
 
-
*This part, BBa_K1017401, includes our artificial sRNA-1 and rRBS-1. The non-coding small RNA can bind to the Shine-Dalgarno sequence on rRBS-1 by base-pairing. Once the rRBS-1 is blocked, ribosomes cannot bind to it to translate, thus, gene expressions downstream are decreased. Because of specific binding, rRBS-1 can only be bound by sRNA-1. We add P<sub>lux</sub> upstream, so this part can be regulated by luxR/AHL  complex. There is one important thing hasn't be mentioned is that it is a temporary sequence which contains the restriction enzyme cutting sites of SpeI, EcoRI and XbaI in order for us to separate them.<br>
 
-
[[File:Nctu_sRNA_rRBS1.jpg|350px]]
 
-
 
-
======BBa_K1017402======
 
-
*This part, BBa_K1017402, is similar to BBa_K1017401 mentioned above, but without P<sub>lux</sub>. rRBS-2 can only be bound by sRNA-2 due to specificity, then gene expressions downstream are decreased. There is one important thing hasn't be mentioned is that it is a temporary sequence which contains the restriction enzyme cutting sites of SpeI, EcoRI and XbaI in order for us to separate them.<br>
 
-
[[File:Nctu_srna2_rRNS.jpg|250px]]
 
  </div>
  </div>

Revision as of 11:19, 10 September 2014

Project

Parts submitted to the Registry

<groupparts>iGEM014 NCTU_Formosa</groupparts>

Brief Information

Please click on the name of the parts for detailed information that is hosted in the Registry website.

PBAN-producted system

PBAN1
  • pcya and ho1 are the enzymes needed to convert heme into chromophore phycocyanobiline(PCB), the red light sensor. Since pcya and ho1 are not naturally produced in E.coli, we use Pcons upstream in order to make E.coli continuously expressed them to synthesize PCB.


450px

PBAN2
  • Cph8 is a chimeric light receptor. It is a fusion of the photoreceptor cph1 and the envZ histidine kinase. cph1 is only active when it binds the chromophore phycocyanobiline (PCB).


150px

PBAN3
  • This is the key part to change Ompc promoter intro red promotor. The lacI will inhibit the function of Plac, thus, the original outcomes will be converted into our expected result.


300px

PBAN4
  • Ompc promtor, which can sense red light with the presence of cph8. It will be turned on in the dark ,and be turned off in the bright. For the convenient use, we add lacI and lac promotor downstream the biobrick. This part, so called red promotor, can be activated under red light, and inactive in the dark.


450px

PBAN5


200px

PBAN6
  • By using mGFP as a reporter gene, we can test whether the 37 °C RBS works.


200px

PBAN7
  • In our circuit, this biobrick is the part of Plux's activation when the temperature reaches to 37oC.


200px


PBAN8
  • The sRNA is the complement of its rRBS. It can regulate the downstream of rRBS in RNA level by binding onto the rRBS when it is transcribed in order to interrupt ribosomes' work. In addition, adding Plux upstream makes the sequence be controlled by luxR/AHL complex.

250px

PBAN9
  • The sRNA is the complement of its rRBS. It can regulate the downstream of rRBS in RNA level by binding onto the rRBS when it is transcribed in order to interrupt ribosomes' work.


200px


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