Difference between revisions of "Part:BBa K2213008"
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Revision as of 19:58, 31 October 2017
HighPromoter_PduD(1-20)_mCherry
This part is an improved version of https://parts.igem.org/Part:BBa_K562001, submitted by Dundee 2011.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 8
Illegal NheI site found at 31 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Function:
This part is an improved version of https://parts.igem.org/Part:BBa_K562001, originally submitted by team Dundee 2011. The original part contained an illegal XbaI site which has been removed to make it biobrick compatible. This part has been expressed under different strength promoters. A high strength Anderson promoter here, low strength Anderson promoter (https://parts.igem.org/Part:BBa_K2213006), and a medium strength Anderson promoter (https://parts.igem.org/Part:BBa_K2213007).
The PduD tag was combined with the high strength Anderson promoter (https://parts.igem.org/Part:BBa_J23104) and mCherry.
A gradient of fluorescence is evident when compared to low and medium promoters.
The expression levels compared to low and medium are as shown.
When expressed alone the distribution of the mCherry signal was homogeneous and fluorescence level was relatively high. In the presence of EutS the mCherry signal was less homogeneous but lacked obvious localisation, suggesting EutS isn’t enough to form proper BMCs. In the presence of EutSMN the mCherry signal clumped together, indicating localisation to the BMC.