Low protein amount - what to do?

Frank Fuerst ffrank at rz.uni-potsdam.de
Wed Feb 28 10:52:30 EST 2001


"Silas Bruun" <sb at licapharma.com> wrote:

>Hi
>I have cloned a gene in the "in vitrogen" pBAD and expressed it in the TOP10
>Coli cells that comes along with the kit. But I really don't get much
>protein (very litle) out of it.
>
>Do anyone know how to boost these cells 

Well, there can be many reasons why the yield is low, ranging from
codon usage (e.g. if it is a eucaryotic protein) or strain-specific
"things" to protein stability and degradation issues. In the latter
cases, you can play with temperature, growth levels upon induction or
even temperature shifts before induction (to induce stress response).

By the way, you've made sure that the protein isn't just in some
insoluble fraction?

Bye, Frank






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