Foreign protein expression in E. coli

Vincenzo Nardi-Dei vinz at PCLSP2.KUICR.KYOTO-U.AC.JP
Sat Feb 5 04:22:09 EST 1994


I have recently cloned an hydrolase of 35,000 M.W. from Pseudomonas
in to E. coli XL1 Blue, provided from Stratagene.
The plasmid used is pUC118, the insert is about 1,200 bp
and the expression is around 10 % of the cell crude extract;
Unfortunately large amount of the enzyme is found in form
of inclusion bodies and the cells take 48 hours (!) to reach
a full growth in LB medium at 37 C and 200 rpm.
This is not the first time I have
inclusion bodies and slow growth using XL1 Blue.
Moreover, a collegue of mine is having the same problem
with the same strain.

In this view I would like to change the host: can anyone
kindly suggest me which strain of E. coli is best fit
for high expression of this kind of foreign proteins?

Thank you very much indeed,

Vincenzo Nardi-Dei

Laboratory of Microbial Biochemistry
Institute for Chemicall Research
Kyoto University
611 Japan




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