Difference between revisions of "Part:BBa K3229304"

Revision as of 16:54, 21 October 2019

McyA/D biderectional promoter 2

This part was constructed to certify the bidirectional transcription from mcyA/D promoter. Therefore we put a composite GFP which coded by two identical genes. Originally the mcyA/D promoter located between the mcyABC and mcyDEFGHIJ, thus the gene expression can occur in both ways. These genes were replaced with the two parts of the composite GFP genes, GFP2 located in the 3' direction and GFP1 is in 5' direction from the promoter sequence. When microcystin toxin is present it induces the synthesis of both of the composite GFP parts due to the phenomenon called autoinduction. The synthesized protein parts bond together and form a functional GFP that glows under UV light. Due to this method, we are able to perceive the appearance of the microcystin toxin and test the bidirectional transcription from the mcyA/D promoter. We can measure the amount of the emitted green light by fluorescence microscope. The composite part consists of a reversed terminator sequence, the reversed sequence of the original GFP1, a reversed RBS, the mcyA/D promoter, another RBS, the GFP2 sequence, a terminator sequence, a noncoding sequence (ATATATATAT) and a BamHI restriction site (GGATCC). We used the reversed sequences of the original sequences we found on the registry pages. These parts were made by GeneiousPrime. 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 both Escherichia coli and Microcystis aeruginosa.

Sequence and Features