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| | <partinfo>BBa_K1660007 short</partinfo> | | <partinfo>BBa_K1660007 short</partinfo> |
| | + | <strong style="font-size: 150%; font-weight: bold;"> (PompC+6N+gp35)</strong> |
| | + | <br/> |
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| | + | <h2>Introduction</h2> |
| | + | 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. |
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| | + | <h2>Design</h2> |
| | + | We insert PompC([https://parts.igem.org/Part:BBa_R0082 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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| | + | 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. |
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| − | PompR 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.
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| − | We insert PompR(BBa_R0082) sequences 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 in through the control of light.
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| − | Bxb1 gp35 is a serine integrase in Mycobacterium phage Bxb1. This intergrase can exclusively catalyze site-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) 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.
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| − | We insert the gp35 gene into the downstream region of the PompR promotor by means of 3A synthesis so that the expression of integrase can be regulated by light. In addition, we add a 6N promotor between PompR and gp35 to decrease basal integrase expression above flipping threshold level which could lower the efficiency of int translation. Meanwhile, we insert the recombined sites of attB and attP into the flanking sequences of our constitutive promotor (BBa_J23110 ) and so far we accomplish our reverse promotor system.
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| | + | </body> |
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| | <!-- Add more about the biology of this part here | | <!-- Add more about the biology of this part here |
| | ===Usage and Biology=== | | ===Usage and Biology=== |
Revision as of 15:36, 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 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.
