Fusion protein and Enterokinase

Lauge Schaffer lsc at novo.dk
Wed Apr 30 08:44:17 EST 1997


Huang Ke-xue wrote:
> Hi, Netters,
> I am experessing some genes in E. coli by using pET32, it gave me good
> result. But the painful problem is when I tried to relase the native protein
> from fusion protein using enterokinase, I could not get the amount what I
> want although I followed the protocol Novagen provided. Sometimes I found
> the protein were degraded (by SDS-PAGE).
> Is it a usual problem for fusion protein or my hands are two bad?
> Huang

Enterokinase is a lousy enzyme (both cloned and natural).
It will cut other sequences with just one or two acidic residues before
the basic one, if they happen to be more accessible than the desired
site.

Lauge



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