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Revision as of 11:51, 20 October 2019

Native pFadD promoter regulating downstream RFP

Description

It is pointed out by the previous iGEM team, UPF CRG Barcelona_2018 iGEM, that the native promoter pFadBA (BBa_K817002) has great leakage and is a weak promoter that isn’t ideal enough for our system. Therefore, to enhance the sensitivity and reduce the leakage of the promoter pFadBA for making it more suitable for our system, we modified the native promoter pFadBA by change consensus sequence of -10 and -35 region. According to the previous research, there is improvement in expression for pFadBA with consensus sequence.

Mechanism

Some study shows that in E. coli, there is sequence called consensus sequence which is universal and change the expression level of downstream protein. Consensus sequence is existing in -10 and -35 region, so it is relative with gene expression.

This picture shows the consensus sequence in E coli and some mutant sequence of this region.


The expression level can be change if sequence is modified. The patterns of the data points indicate which promoter clones contained mutant in either the -35 or the -10 region with consensus sequence.


Usage and Biology

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Unknown
  • 23
    INCOMPATIBLE WITH RFC[23]
    Unknown
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 755
    Illegal AgeI site found at 867
  • 1000
    COMPATIBLE WITH RFC[1000]


Functional Parameters