Any experiences with pET 31b(+) Vector and KSI-Fragment purification?

Leif Dehmelt leif.dehmelt at mpi-dortmund.mpg.de
Fri Jul 4 06:06:53 EST 1997


Hi all!

I would like to read about some good or bad experiences
with the pET 31b(+) Vector from NOVAGEN. This Vector decodes 
for a KSI-Fragment Fusion with C-terminal His-Tag. The KSI
(Ketosteroid-Isomerase)-Fragment leads to insolubility and strong 
overexpression in inclusion bodies, even for small proteins or peptides
in fusion (17-65 as). The KSI-Fusion is cleaved by CNBr and the following 
seperation step is achieved by simple centrifugation.

Does anybody have particular practical experiences with this system?

Thanks for every reply...

Leif



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