pET, lysE, lysS, bacterial expression questions

jfh jfhess at ucdavis.edu
Thu Jun 2 12:00:51 EST 1994


Howdy gang,

I have experience using pT7-7 (precursor to pET) for expression of
proteins in bacteria. However, our labs latest expression attempts (our
largest protein so far, also, 86 kD) are less than stellar.  we went from
BL21 (DE3) to the lysS and lysE strains to help.

Our major problems are 1) expression of the protein in UNinduced cells
and 2) not major increases in exression upon induction. 

My questions are 1) any better strains to try with my existing pT7
construct?
2) anyone have experience using rifampicin to shut down the e colis?
3) anything else to try?


John Hess, PhD                    Phone me 916 752 8420
Dept of Human Anatomy             FAX me 916 752 8520
University of Calif               Email me jfhess at ucdavis.edu
Davis, CA                         or leave me alone, your choice.



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