Difference between revisions of "Part:BBa K3040005"
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Revision as of 11:56, 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.
![](https://static.igem.org/mediawiki/parts/0/05/T--NTHU_Taiwan--liya1.png)
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.
![](https://static.igem.org/mediawiki/parts/b/b0/T--NTHU_Taiwan--liya2.png)
Method
1. Clone this promoter into DH5a
2. Detection fluorescence level after adding different concentration of fatty acid.
3. Analysis the data
Result
We detect the fluorescence by the mechanism. The following is the picture.
Usage and Biology
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 755
Illegal AgeI site found at 867 - 1000COMPATIBLE WITH RFC[1000]