Difference between revisions of "Part:BBa K4242017"

Revision as of 12:18, 12 October 2022

Ptac+RBS+yhjH+TT（pSB3C5-A3）

Uses promoter Ptac(BBa_K180000), RBS(BBa_J34801), protein YhjH coding region(BBa_K861090) and TT(BBa_B0015). This part is a device to construct the expression of protein YhjH that waas controlled by the IPTG inducible promotor Ptac

Usage and Biology

This part is improved from BBa_K2471001to express the c-di-GMP hydrolase YhjH. With the expression of this gene, intracellular c-di-GMP is reduced, causing the decrease of biofilm. However, when we test its function in Shewanella oneidensis MR-1 strain, we found the biofilm formation didn’t decrease compared to the control group. This is because S. oneidensis strain doesn’t contain the T7 RNA polymerase needed for the T7 promotor. So we improved the original part and created this part by switching the consititutive promotor T7 to the IPTG inducible promotor Ptac. In this way we can not only control the yhjH expression level by adding IPTG, but also make it functional in a wider variety of strains who lack the T7 expression systems. We measured biofilm by the method of crystal violet staining, as the result shown in Fig.1, biofilm formation was significantly reduced with the addition of inducer IPTG compared with the control group, indicating this new part is well functional in S. oneidensis MR-1 strain.

fig.1 a|Structure of control biobrick pSB3C5-Ptac b|Structure of pSB3C5-T7-YhjH BBa_K2471001 c|Structure of improved part pSB3C5-YhjH BBa_K4242017

Sequence and Features