Dear Colleagues, I am currently trying to overexpress RNAP(B.subtilis) in E.coli strain B834 (DE3) pLysS. My plasmid DNA(pNG219) contains three of RNAP subunit genes which are downstream of T7 promoter. I do fresh transformation but still get very low amount of RNAP(with 9x his-tag). Instead of RNAP but Beta-galactosidase is overexpressed. Does anybody have this problem? I followed protocol for pNG219 that has shown below. http://www.ncbi.nlm.nih.gov/pubmed/18289874 Thanks, Hun _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
