Difference between revisions of "Part:BBa K2253002"

Revision as of 01:04, 1 November 2017

Optimized Blue Chromoprotein

This is the original blue chromoprotein sequence (BBa_K592009) that has been codon optimized. The original part has been made a basic part with the promoter and RBS sequence, BBa_K608002 in the pSB1C3 BioBrick backbone. Codon optimization allows it to function more efficiently in E.coli, therefore creating a lower metabolic burden on the cells that it is encoded in. When this sequence is put into a vector and transformed into a plasmid it shows a blue phenotype. Codon optimization also makes this part more stable, as the color is expected to be maintained for a longer period of time than the original, making it a good candidate to use as a biosensor for things like toxins, such as lead.

Experimental Design

To attempt to improve the stability of the original blue chromoprotein, a digestion was performed on the K608002 vector and the codon optimized blue chromoprotein, which was ordered as a G-block from the Integrated DNA Technologies (IDT) website. Upon digesting each of these parts and conducting a gel extraction, a ligation reaction was used to ligate the gene of interest (codon optimized blue chromoprotein) into the K608002 vector. After purifying this ligation, a transformation was performed, which yielded in one phentotypically blue colony, shown below.

Figure 1. Transformation plate of the K60800 vector and codon optimized blue chromoprotein ligation reaction.

Sequencing Results

Through the codon optimization of this part, in comparison to the BBa_K592009 original chromoprotein, the sequenced codon optimized blue chromoprotein shows minimal mutations.