Team:Tianjin/Judging

BBa_K1361000

Based on the concept that change of cell motility by control of CheZ chemotaxis regulator could disturb pre-expressed curli fiber structure, we assume that if the inducer occurs, in this case acacia, CsgB protein will secrete into extracellular and act as nucleator to recruit soluble CsgA monomer into amyloid fibres, and the transcription of CheZ will be downgrade via cI-Plamda NOT gate. If induction condition disappears at a certain point, suppression of CheZ will be discharged. As a result, cells will be more moveable thus disturb or inhibit the establishment of curli structure. And in our final device, the dynamic detection could achieve by monitoring current change between two electrodes in the culture.

BBa_K1361001

This part contains the major subunit of curli fiber CsgA and its nucleator CsgB, in which CsgA is constitutive expression(a relatively weak promoter compared to BBa_K1361003) whereas CsgB is under the control of T7 promotor. This part cannot functioning alone without expression of CsgEFG genes in the same cell, for they coding the outer membrane channel secrete system for CsgA and CsgB.

BBa_K1361002

This part contains the major subunit of curli fiber CsgA and its nucleator CsgBtrunc, in which CsgA is constitutive expression whereas CsgBtrunc is under the control of Pbad promotor. CsgBtrunc is different from native CsgB for the deletion of C-terminal amino acid from 133 to 155 so that it cannot attach cell outer membrane. And CsgBtrunc itself is highly aggregative in the culture. This part is designed for immediate generation of large amount of curli fiber after adding the inducer. This part cannot function alone without the expression of CsgEFG genes in the same cell, for they coding the outer membrane channel secrete system for CsgA and CsgB.

BBa_K1361003

This part contains the major subunit of curli fiber CsgA and its nucleator CsgB, in which CsgA is constitutive expression(a relatively strong promoter compared to BBa_K1361001) whereas CsgB is under the control of T7 promoter. This part cannot function alone without expression of CsgEFG genes in the same cell, for they coding the outer membrane channel secrete system for CsgA and CsgB.

BBa_K1361004

This part contains the major subunit of curli fiber CsgA-his (CsgA modified by His tag) and its nucleator CsgB, in which CsgA-his is constitutive expression whereas CsgB is under the control of Pbad promoter. This part is designed for immediate generation of large amount of curli fiber after the adding of the inducer and the purification of CsgA by Ni-NTA reign or absorbing Au-NTA-Ni nano partial. This part cannot function alone without expression of CsgEFG genes in the same cell, for they coding the outer membrane channel secrete system for CsgA and CsgB.

BBa_K1361005

This part contains CsgE, CsgF, CsgG, which serve as the outer membrane secrete device for curli fiber, at relatively low constitutive. This part is for specific transport of CsgA(BBa_K1361999) and CsgB(BBa_K1361997) into extracellular matrix. CsgEFG(BBa_K1361992) was placed after a relatively weak constitutive promoter.

BBa_K1361006

This part contains the major subunit of curli fiber CsgA and its nucleator CsgBtrunc (a dissociative nucleator), in which CsgA is constitutive expression whereas CsgBtrunc is under the control of Pbad promoter. CsgBtrunc is different from native CsgB for the deletion of C-terminal amino acid from 133 to 155 so that it cannot attach cell outer membrane. And CsgBtrunc itself is highly aggregative in the culture. This part is designed for immediate generation of large amount of curli fiber after the adding of the inducer, and a stronger promoter for CsgA was chosen to parallel with BBa_K1361002. This part cannot function alone without expression of CsgEFG genes in the same cell, for they coding the outer membrane channel secrete system for CsgA and CsgB.

BBa_K1361007

This part contains the major subunit of curli fiber CsgA-his CsgA modified by His tag) and its nucleator CsgB, in which CsgA-his is constitutive expression whereas CsgB is under the control of Pbad promoter. This part is designed for immediate generation of large amount of curli fiber after adding of the inducer and the purification of CsgA by Ni-NTA reign, or absorbing Au-NTA-Ni nano partical. This part cannot function alone without expression of CsgEFG genes in the same cell, for they coding the outer membrane channel secrete system for CsgA and CsgB.

Requirements for the Bronze and Silver Medal:

√Register the team, have a great summer, and plan to have fun at the Giant Jamboree.

√Successfully complete and submit this iGEM 2014 Judging form.

√Create and share a Description of the team's project using the iGEM wiki and the team's parts using the Registry of Standard Biological Parts.

√Plan to present a Poster and Talk at the iGEM Jamboree.

√The description of each project have clearly attribute work done by the us and distinguish it from work done by others, including host labs, advisors, instructors, sponsors, professional website designers, artists, and commercial services.

√Document more than one new standard BioBrick Part or Device central to our project and submit these parts to the iGEM Registry (submissions must adhere to the iGEM Registry guidelines).

√Experimentally validate that new BioBrick Part or Device of our own design and construction works as expected.

√Document the characterization of new parts in the Main Page section of that Part's/Device's Registry entry.

√Submit this new part to the iGEM Parts Registry (submissions have been adhere to the iGEM Registry guidelines)

√We carried out several human practice activities, such as a visit to TIB,CAS (Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences), a regular newsletter with more than 20 iGEM teams all over the world and a nationwide questionnaire focus on the spread of synthetic biology. Besides, we have done mutual help with NKU and BIT iGEM teams in terms of experimental instructions and future applications.

Additional Requirements for a Gold Medal:

√We have helped Beijing Institute of Technology( BIT ) team to model their MIN system, using a classical density-dependent growth model for the population of E.coli growing simulation. Also we use some other differential equations to describe the dynamic change of AHL, LuxR proteins and MinC proteins. Apart from that, we helped Xiamen University (XMU) team start the regular newsletters with over 20 teams around the world.

√Our project has the implications for the nanowire devices focus on the bio-signal transformation. Our project provides a novel and controllable method to diversify the output methods of biosensor, though at a preliminary stage. With future work, a more stable, sensitive and economical device should be promising.

Tianjin University,Tianjin, China
Email:michaelss@tju.edu.cn