Difference between revisions of "Part:BBa K4779002"

Revision as of 19:15, 11 October 2023

CMPS:A yeast copper-induced reporter pathway with Positive feedback loop in with prm1 promoter

It is a copper ion reaction reporting pathway in Saccharomyces cerevisiae. We employed a positive feedback regulation with Ste5ΔN-CTM for signal amplification. In addition, we utilized the prm1 promoter for both GFP and Ste5ΔN-CTM expression.

Construction

CMPS is a new composite part (BBa_K4779002). We obtained the gene sequences of the promoter of Cup1 (BBa_K945002), MFα2 (BBa_K110005), the promoter of prm1 (BBa_K1346004), green fluorescent protein (GFP) (BBa_K3112009), Ste5ΔN-CTM (BBa_K3384315) and the terminator CYC1 (BBa_K4278703) from IGEM, and sent them to GENEWIZ (Suzhou Genewiz Biotechnology Co., Ltd.) for synthesis.

Characterization

We subjected the engineered yeast strain BY4741-pRS415-CMPS samples to a copper ion gradient treatment and quantitatively measured their fluorescence intensity at different copper ion concentrations, as depicted in the graph. Within 500 uM, the signal output intensity of the CMPS sensor increased with an ascending copper ion concentration and nearly reached its peak at 50 uM. In comparison to CMPG, which reached its signal output peak at 200 uM, CMPS achieved comparable signal output intensity at lower copper ion concentrations, indicating an improved sensitivity.

Sequence and Features