发信人: justsososo (justsososo), 信区: Biology
标 题: Re: recombinant蛋白质表达问题
发信站: Unknown Space - 未名空间 (Sat May 1 16:37:48 2004) WWW-POST

BL21codon+ is indeed BL21 with a plasmid that expresses rare codons and
BL21(DE3)pLysS/E is indeed BL21 with a plasmid that expresses an enzyme that
degrade T7 polymerase to avoid leaky. Both plasmid are chlorepenicol
resistance. In inder to make codon plus in BL21(DE3)pLysS/E strain, drug
marker of one plasmid must be change and the system will become too
complicated - the strain resist to two drug, plus another drug marker for your
recombinant protein.

【 在 arabidopsis (moon) 的大作中提到: 】
: I think it doesn't matter if it's leaky or not. Just feel courious about
it.
: Then I wonder why they do not make codon plus in BL21(DE3)pLysS/E strain.
:
: 【 在 justsososo (justsososo) 的大作中提到: 】
: : Yes. Not only BL21codon plus, but almost all BL21s except BL21(DE3)pLysS/E
are
: : leaky. pLysS/E encode enzyme that degrade T7 polymerase to avoid leaky.
: : In your case with many rare Arg codons, you have to use codon plus. So, it
is
: : importment to avoid leaky? otherwise, just express with leaky if it does
no
: : harm.
: : 【 在 arabidopsis (moon) 的大作中提到: 】
: : : I tried this strain and cannot get my protein induced. There are lots
of
: : : rare Arg codons in my DNA sequence. That's why I chose BL21 codon plus.
: : : BL21 codon plus usually leaky?
:
:


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