Team:Peking/Notebook

• Drylab
• Wetlab

Wetlab

Junyi Jiao

It is highly expected that when fused with N-ternimnal fragment of Outer membrane protein A (OmpA), the mercury metal binding peptide will be translocated and displayed on the surface of bacteria. In our design, I conducted this experiment.

Ying Sheng

My work mainly focuses on the characterization of merOP. The response curve of merOP was characterized in Ecoli strains possessing different genetic component combinations to figure out the affects of different module to merOP and finally contributed to the rational design of the bioreporter.

Yang Hu

For the bench work, I was responsible for the construction of mercury metal binding peptide cytosol expression module and T7 promoter drove MerR generation module. I also helped to conduct western blotting assay.On the other hand, I take charge of the work of Human Practice concerning about the potential problems of horizontal gene transfer.

Donghai Liang

Being responsible for the periplasmic module of the bioabsorbent display, I successfully completed the construction of DsbA-MBP and DsbA-MerR (serving as a control). At the same time, the standardization of the module mentioned above is also completed. Besides, I participate in the TF-DNA binding affinity characterization of the mercury promoter.

Yiwei Chen

I expanded the MerR protein engineering strategy to lead-responsive regulator, PbrR. Outer membrane protein A (OmpA) was fused with Pb binding peptide at its C-terminal via the same method in mercury binding peptide construction. I also participated the construction of mercury absorption facilitation module and inductive aggregation module.

Ao Liu

I Conducted mutagenesis library for Mer OP and research on the feature of promoter functional sites through screening the library (characterizging promoters with different dyad MerR binding site). Then promoters with different Hg binding affinity was integrated with the results gained from operation analysis to acquire the wider range of bioreproter Hg-sensitivity.

Boxuan Zhao

My work is merging OmpA and anchor protein Lpp with metal binding domain of MerR to accomplish the goal of surface display. Additionally, I participate the functional testing of whole-cell bioabsorbents. By recognizing the homologous regions among them, I designed the metal binding domain complex for four different MerR family proteins, which can be used to verify the reasonableness of our project design.

Qianzhu Wu

To characterize the regulating role of MerR on PmerT, promoters from parts-registry constutitive promoter library with different strength were prefixed before MerR coding sequence to exogenously maintain MerR expression at different intensity. GFP expression response to mercury concentrations was measured by microplate reader.

Xin Teng

After carefully studied the structure of MerR, I constructed MerR-MBP (Metal Binding Peptide). Additionaly, I helped to construct and standardize the periplasmic metal binding peptide – DsbA-MerR-MBP using a similar strategy as MerR-MBP. After inductive expression of MerR-MBP-His6, DsbA-PbrR-MBP-His6 and DsbA-MerR-MBP-His6, I use SDS-PAGE and western blot to confirm the expression and localization of MBP and DsbA-MBP.

Mei Chen

I characterized the CrtEBI (lycopene gene) biobrick K274100 submitted by team Cambridge 2009, for which two new biobricks was constructed. Addtionally, I contributed to the bioreporters partly, such as bioreporters for mercury or lead.

Zairan Liu

I am responsible for the construction of metal binding peptide periplasm display module for Pb(II). This module aims at binding Pb(II) ions in the periplasmic space using the engineered anti-parallel coiled coil which is transported with the help of DsbA signal sequence. During the process several other intermediate plasmids are also constructed. Furthermore, I contribute to the characterization of Pc promoters of different intensity.

Heng Pan

I have constructed a genetic circuit, which utilizes the T3 polymerase/ T3 promoter and phage activator PhiR73/PO pomoter pairs to achieve a time dlay and signal amplification after mercury is present.Besides, I have characterized PpbrA-PbrR pair to stress that the engineering strategy applied in PmerT-MerR pair is robust for MerR family TFs

Haoqian Zhang

I have played several roles during this summer. As a team leader, I presided designing and optimization of the project. I know every member of Peking iGEM team is extraordinarily talented and enthusiastic at synthetic biology, which means that all they need is proper coordination and guide. Therefore, I did not join the bioware experiments very much, but worked as both a technician and an advisor.

Miao Jing

I designed to engineer this operon under the regulation of T7 promoter, which is regulated by upstream T7 polymerase, to fulfill the goal that the absorbance of Hg(II) will be enhanced for an efficient bioabsorbent. Also I cloned and assembled agn43 into the inductive aggregation module. Agn43 is drove by PO promoter, which is the terminal of a cascade amplification

Weiye Wang

My wetlab mission is to select a T3 promoter with medium expression level from T3 phage genome: Constructing 3 multi-plasmid systems, using IPTG, mercury and arabinose as the inducer of T3 polymerase’s expression, so that to characterize the response properties of these T3 polymerase dependent promoters. The design and construction of our team's wiki is also under my charge.

Tianze Zhu

Drylab

Zhenzhen Yin & Yuheng Lu

Using computional methods to search for proper topology whose input and output have an linear correlationship. Additionally They conduct modeling to explain and simulate the operation mechanism of MerR family TFs on their cognate promoters.