Difference between revisions of "Part:BBa K3229302"

Revision as of 16:14, 21 October 2019

McyA promoter reporter (with reverse promoter)

This part is appropriate to measure the activity level of the putative mcyA promoter region (reversed mcyD). Normally the promoter is next to the genes of microcystin toxin-producing proteins (mcyABC and mcyDEFGHIJ) in both 5'-3' and 3'-5' directions. We replaced these genes with a GFP firstly in 5'-3' direction. Only if the toxin microcystin present in the sample, the toxin induces the expression of the GFP gene due to the putative promoter sequence. As a result, the GFP glows under UV and we can measure the intensity of the emitted light, by fluorescence microscope. According to this, the emitted light correlates with the amount of toxin present. This composite part consists of the promoter sequence (mentioned above), an RBS, a GFP, a terminator sequence (this three was selected from the registry), a non-coding sequence (ATATATATAT), and a BamHI restriction site (GGATCC). The uploaded part does NOT contain the non-coding and the BamHI sequence. The non-coding sequence and the BamHI are required to insert the construct into a shuttle vector which we can transform the plasmid into Microcystis aeruginosa.

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Sequence and Features