Yoram Gerchman (gerchman at Princeton.EDU) wrote:
> Greetings all
> I want to express an high activity alkaline phosphatase in
> E. coli. Anyone has an idea for high activity AP gene that
> work in E. coli? I know Roche got a recombinant version of the
> bovine AP but I could not find any reference as to how it was done.
> Thanks Yoram
> -------------------------------------------------------
> "Support bacteria, it's the only culture some people have."
> Anonymous
>
I would suggest using the online genome searching resources. Find the
bovine AP gene, get some gDNA, clone the gene out, insert it into a
bacterial vector, express it, purify it, and test it. If that doesn't
yeild useable protein, try using a eukaryotic expression system. I like
the high 5 insect cell line. It has the best qualities of adherent and
suspended cultures all in one.
