Difference between revisions of "Part:BBa K1660007"
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| − | We insert PompC([https://parts.igem.org/Part: | + | We insert PompC([https://parts.igem.org/Part:BBa_R0082]) sequence into the upstream region of our target gene gp35 through restriction-ligation method, which together is connected to vector pSB1C3 afterwards, and therefore we are able to regulate the expression of int through the control of light. |
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Revision as of 15:38, 17 September 2015
PompR-6N-gp35-Terminator
(PompC+6N+gp35)
Introduction
PompC is a OmpR-controlled promoter which can be positively regulated by phosphorylated OmpR. This promoter is taken from the upstream region of ompC. Phosphorylated OmpR binds to the three operator sites and activates transcription.Bxb1 gp35 is a serine integrase in Mycobacterium phage Bxb1. This intergrase can exclusively catalyzesite-specific recombination. The dismiss of light singal can drive the expression of integrase, which alone can set a DNA sequence (there it means the promoter BBa_J23110) flanked by oppositional attB and attP sites, thereby produce an inverted sequence flanked by attL and attR sites. As a consequence, the promotor moves on to express the toxic protein instead of the attractant. We connected PompC and gp35 together to form this part whose length is 1773bp.
Design
We insert PompC([https://parts.igem.org/Part:BBa_R0082]) sequence into the upstream region of our target gene gp35 through restriction-ligation method, which together is connected to vector pSB1C3 afterwards, and therefore we are able to regulate the expression of int through the control of light.In addition, we add a 6N promotor between PompC and gp35 to decrease basal integrase expression above flipping threshold level which could lower the efficiency of int translation.