Generator
Part:BBa_K223062:Experience
Designed by: Anusuya Ramasubramanian Group: iGEM09_Stanford (2009-10-18)
Revision as of 15:22, 19 October 2011 by Favaro.mtp (Talk | contribs)
New Secretion System by [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil UNICAMP-EMSE] 2011 Team based on Stanford's 2009 System
This part device has been improved by UNICAMP_EMSE team (2011) and is now functional. Improvements include:
- Change the RBS part and its disposition in the transcriptional unit.
- Attempting to create a new secretion system, we began by choosing a different but still highly efficient RBS part. And according to the experience made upon RBS deposited parts (BBa_B0034), it was suggested that the RBS chosen by our team (B0030) is as efficient as the one used by Stanford 2009 team (B0034).
- To improve translation efficiency, gene sequence fidelity (avoid errors or mistranscription during PCR) and save time during the devices assembling UNICAMP-EMSE team synthesized each coding sequence already fusioned with RBS sequence. On the other hand Stanford 2009 Team did not use translational units.
- 2. Biobrick Standard Compatibility
- The secretion system developed by Stanford Team 2009 was not compatible with the systems 10, 12, 21, 23 nor 25. UNICAMP-EMSE Team has removed internal illegal restriction sites presented in the parts Bba_K223055 and BBa_K223057 by codon usage of these sites prior to synthesis. Therefore our team suited the secretion system to be compatible with 10, 12 and 23 standards.
- 3. Exclusion of non-essential genes
- In our system we have reduced the number of genes to use only the essential ones, considering that “the simpler, the better”. Evidence [] shows that only TolC, HlyB and HlyD are essential for the system to work, hence we have excluded the other genes used by Stanford 2009 Team (Part Bba_K223056).
- 4. HlyB and HlyD
- We have opted not to use HlyB and HlyD as a fusioned unit, like Stanford 2009 Team did (Part BBa_K223057), but as two independent translational units (Part BBa_K554008 and Part BBa_K554007)
- See additional details, including functional tests with GFP, on the [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Results results] section of our wiki.
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UNIQ6e3f457a3fb1ea88-partinfo-00000000-QINU UNIQ6e3f457a3fb1ea88-partinfo-00000001-QINU