Part:BBa_K3786004
TMM expression controlled by the Arabinose promoter
When transformed into E. Coli, this part allows for the production of TMM (Trimethylamine moonoxygenaseonly) when there is presence of Arabinose and low glucose levels in the medium. The protein of interest needs to be located downstream of the operon to be induced by arabinose. This part contains the pBAD promoter, B0030 ribosome binding site, TMM gene sequence and B0010 terminator. The TMM expression cassette transforms trimethylamine into trimethylamin-N-oxide and can also lead to the production the blue pigment Indigo. Therefore, this part allows to have inducible Indigo production in the presence arabinose .
Results and Characterization
We transfomed this plasmid into TOP10 cells for transformation efficiency. We can see a difference in color between the cells transformed with our TMM plasmid and cells not containing the plasmid as shown in Figure 1. The cells containg the plasmid a darker, almost orange. This shows the production of indoxyl, the precursor of indigo. The plasmid is a medium-copy plasmid. This can explain why the color is not very strong. Further characterization will be conducted to confirm and quantify the expression of TMM by the cells. (Fig. 1).
Figure 1: Picture of colonies showing a comparison of NEB Top10 cells transformed with the pTS000 plasmid containing TMM and wild-type E. coli.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 239
Illegal BamHI site found at 663 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 74
Illegal AgeI site found at 487
Illegal AgeI site found at 710 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 56
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