Difference between revisions of "Part:BBa K2213006"

Revision as of 01:30, 1 November 2017

LowPromoter_PduD(1-20)_mCherry

This part is an improved version of https://parts.igem.org/Part:BBa_K562001, submitted by Dundee 2011.

The PduD(1:20) tag is one of many tags that has been proved to localise inside a Pdu microcompartment. This has been proved by the iGEM Dundee 2011 team http://2011.igem.org/Team:Dundee. This part has been tagged with the fluorescent protein, mCherry, to facilitate its characterisation, as can be seen in the figure below:

figure1. circuit diagram of BBa_K2213006.

Improvements and Characterisation

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 also been expressed under different strength promoters.
A low strength Anderson promoter (here)
medium strength Anderson promoter (https://parts.igem.org/Part:BBa_K2213007)
high strength Anderson promoter (https://parts.igem.org/Part:BBa_K2213008)

The PduD tag was combined with the low strength Anderson promoter (https://parts.igem.org/Part:BBa_J23105) and mCherry.



A gradient of fluorescence is evident when compared to medium and high promoters.




The expression levels compared to medium and high are as shown.





When expressed alone the distribution of the mCherry signal was homogeneous and fluorescence level was very low making visualisation difficult. In the presence of EutSMNLK the mCherry signal was much brighter and clumped together, indicating localisation to the BMC.

Sequence and Features