No RNAP but beta-galactosidase

hanna jangg via methods%40net.bio.net (by hunnajangg from hotmail.com)
Fri Nov 19 10:45:38 EST 2010


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  		 	   		  


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