Team:SJTU-BioX-Shanghai/Project/Light sensor/Green
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Certain sgRNA is under control of the green light sensing system. sgRNA targeting RFP is cloned and the base pairing region was easily replaced by reverse PCR to target luciferase and FebI. | Certain sgRNA is under control of the green light sensing system. sgRNA targeting RFP is cloned and the base pairing region was easily replaced by reverse PCR to target luciferase and FebI. | ||
- | But there is a problem we | + | But there is a problem we cannot ignore, sgRNA transcripts must be strictly precise to function normally. Redundant deoxyribonucleoside in 5’-terminal of sgRNA may cause inactivation. Considering that we do not know transcription initiation site of CpcG2 and, more importantly,in order to be consistent with other light sensing systems, we did some modification to CcaR-responding promoter. |
- | Previous studies have shown the G-box of CpcG2 | + | Previous studies have shown phosphorylated CcaR recognizes and binds to the G-box of CpcG2.[2]Six base pairs in the G-box are identical to -35 site of J23107(A constitutive promoter). So we overlap G-box to BBa_J23107. CcaR will bind to G-box, thus prevents RNA polymerase from binding to the promoter. This modification(BBa_K1023009) allows down-regulation effect under green light. |
[[File:13SJTU_green_design.png|thumb|500px|center|''Fig.1'' BBa_K1023009 design]] | [[File:13SJTU_green_design.png|thumb|500px|center|''Fig.1'' BBa_K1023009 design]] |
Revision as of 01:43, 28 September 2013
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