Agrochemical control of gene expression using evolved split RNA polymerase. II

Abstract

Agrochemical inducible gene expression system provides cost-effective and orthogonal control of energy and information flow in bacterial cells. However, the previous version of Mandipropamid inducible gene expression system (Mandi-T7) became constitutively active at room temperature. We moved the split site of the eRNAP from position LYS179 to position ILE109. This new eRNAP showed proximity dependence at 23°C, but not at 37°C. We built Mandi-T7-v2 system based on the new eRNAP and it worked in bothEscherichia coli and Agrobacteria tumefaciens. The modified eRNAP when combined with the leucine zipper-based dimerization system, behaved as a cold inducible gene expression system. Our new system provides a means to broaden the application of agrochemicals for both research and agricultural application.