[Microbiology] Re: Expressing high activity AP in coli

Mike McWilliams michael.mcwilliams at drdc-rddc.gc.ca
Wed Jul 6 13:19:58 EST 2005


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.


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