Regulatory

Part:BBa_K4156106

Designed by: Zheng Huang   Group: iGEM22_LZU-CHINA   (2022-10-07)
Revision as of 14:21, 10 October 2022 by Liwei (Talk | contribs)


pPepT-Bxb1

pPepT-Bxb1 is constructed with hypoxia-sensing promoter pPepT and serine integrase Bxb1.


Usage and Biology

pPepT-Bxb1 consists of a fusion of the pH-sensitive promoter pPepT and the serine integrase Bxb1. It will act as a complex regulatory for controlling downstream logic gates and transcription of genes.

Characterization

In vitro characterization and data analysis of the reported strains

To improve signaling stability as well as accuracy, we added Amplifying genetic switches based on serine integrase (Bxb1,TP901) to the R reporter(BBa_ ) to construct the AR reporter. Fig 1 indicates hypoxia (pPepT）induced AR reporters with homogenized fluorescence intensity (mRFP/Cell). Comparing Fig.1 and 2, the fluorescence expression of the AR reporter was significantly higher after the addition of Switch under anaerobic conditions. This result indicates that the addition of amplifying genetic switch enhances the reporter intensity and robustness of the lactate biosensor.

control
Figure 1: Induction of downstream gene mRFP expression over time by the AR reporter consisting of pPepT+Switch+mRFP under hypoxic and normoxic conditions.

control
Figure 2: Induction of downstream gene mRFP expression over time by the AR reporter consisting of pPepT +mRFP under hypoxic and normoxic conditions.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 360
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 634
    Illegal XhoI site found at 721
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1468


[edit]
Categories
Parameters
None