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Revision as of 10:56, 10 September 2015
Extraction of gene fragements
We successfully get gene fragements
that are used for bacterial permeability improvement, which including,
micF and SoxS, these two antibiotic substance responding promoters were extracted from E. coli Top 10(K-12 strain) by conducting polymerase chain reaction on its genome.
SCVE, a viroporin originally from SARS virus, is synthesized from Sangon Co. ctd.
- OprF, a bigger porin compared with OmpF in E. coli, is extracted from P.aeruginosa,
PAO1 genome by PCR.
- CheZ, a chemotaxis related protein is also extracted from P.aeruginosa, PAO1 genome by PCR.
- Cas9, from 2015 part registry.
- gRNA-AcrB and gRNA-EmrE, these two fragments is for genetically modifed bacterial
transmembrane protein AcrB and EmrE to strongly block drug efflux system.
Growth characteristics
We detected OD600(nm) to identify the effect
of cheZ expression on E. coli through time.
The exogenous protein expression affects the growth of bacteria, which was indicated by slight slower grow-up in E. coli containing cheZ plasmid. In total, cheZ doesn't have obvious negative impact on bacteria grown.
SDS-PAGE
After induced by IPTG in 1mM about 4 h, cheZ was found in
SDS-PAGE gel, compared to the wild type (Top10) and bacteria without IPTG induction.
Through this result, we concluded that cheZ has been successfully expressed with the
IPTG induction.
See
protocol in Protocols: SDS-PAGE
Code Name | Parts Name | Type | Function |
---|---|---|---|
micF | Simple/Promoter | ||
SoxS | Simple/Promoter | ||
OprF | Simple/CDS | ||
SCVE | Simple/CDS | ||
cheZ | Simple/CDS | ||
T7-RBS-SCVE | |||
T7-RBS-OprF | |||
R0010-RBS-cheZ | |||
micF-RBS-GFP | |||
SoxS-RBS-GFP | |||
tRNA&tRNA Synthetase | |||
Compx | |||
tRNA complex with Compx |
Bronze:
Your team must convince the judges you have achieved the
following 6 goals:
- Register for iGEM, have a great summer, and attend the Giant Jamboree.
- Complete the Judging form.
- Create and share a Description of the team's project using the iGEM wiki, and document the team's parts (if any) using the Registry of Standard Biological Parts.
- Present a poster and a talk at the iGEM Jamboree. See the 2015 poster guidelines for more information.
- Create a page on your team wiki with clear attribution of each aspect of your project. This page must clearly attribute work done by the students and distinguish it from work done by others, including host labs, advisors, instructors, sponsors, professional website designers, artists, and commercial services.
- Develop a working prototype of an open-source hardware tool that supports synthetic biologists working with BioBrick standard biological parts. Document your tool on the project page on your team wiki.
Silver:
In addition to the Bronze Medal requirements, your team must
convince the judges you have achieved the following 3 goals:
- Provide a demonstration of your hardware using a video tutorial that explains all the features of the hardware operation. Make the video available on your wiki. You must also send the video file to iGEM HQ (email hq [at] igem [dot] org). See our tutorial video in Tutorial-Instructional Video
- Document your device on your wiki with information and associated files sufficient for a future iGEM team to reproduce your work. You should include relevant design schematics, 3d print files (or other), software and a Bill of Materials (BOM). Want to reproduce our NDM? Please visit Tutorial-Make your own
- iGEM projects involve important questions beyond the bench, for example relating to (but not limited to) ethics, sustainability, social justice, safety, security, and intellectual property rights. We refer to these activities as Human Practices in iGEM. Demonstrate how your team has identified, investigated and addressed one or more of these issues in the context of your project. (See the Human Practices Hub for more information.) See our Practices in Policy&Practices
Gold:
In addition to the Bronze and Silver Medal requirements, your
team must convince the judges you have achieved at least two of the following goals:
- Choose one of these two options: (1) Expand on your silver medal Human Practices activity by demonstrating how you have integrated the investigated issues into the design and/or execution of your project. OR (2) Demonstrate an innovative Human Practices activity that relates to your project (this typically involves educational, public engagement, and/or public perception activities; see the Human Practices Hub for information and examples of innovative activities from previous teams).See our Integrated Practices Activities in Policy&Practices-Integrated Human Practice
- Help any other iGEM team utilize the hardware developed by your team. You must clearly show how your hardware was used and the results that were obtained. Results should take the form of a written report from the team testing the hardware and be posted on both team's wikis.
- Select a previous iGEM Hardware project from any track and demonstrate how you have continued to work on the project by improving the functionality, ease of use, or documentation of another facet of the work/device.
- Demonstrate your hardware at the iGEM Giant Jamboree. If your equipment is too large to transport, create an installation to demonstrate functionality. Note: you may not bring live biological materials to the Jamboree.