Get problems in overexpression protein in E.coli

Duncan Clark junk at hgmp.mrc.ac.uk
Sat Apr 5 05:20:54 EST 2003


Historians believe that in newspost 
<20030401202908.16981.qmail at ww02.hostica.com> on Tue, 1 Apr 2003, 
yourslynn at hotmail.com penned the following literary masterpiece:
>Hellow!I am overexpressing GST-fused protein in PET-41a vector in BL21(pLysS)host strain now.I have totally five proteins for overexpression.But
>only two get overexpressed.I tried the different incubation temperature and time,and with detergent addition on the other three,but failed.I have tested
>the plasmid stability,and find it's stable.So what's going on in my work?

Presumably you have sequenced and confirmed that what you have is 
correct?

What is the source of the proteins, eubacterial, mammalian etc.?

Inclusion bodies?

Could the ones not expressing be using rare codons by E.coli codon 
usage?

Duncan
-- 
I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.



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