in vitro alpha complementation

John Rebers jrebers at nmu.edu
Mon Aug 2 17:53:57 EST 1999


I would like to set up an in vitro alpha complementation assay, mixing a
fusing protein made using the alpha fragment of lacZ with a cell-free
extract containing an alpha-donor (aka omega) fragment, and then assaying
for beta-galactosidase activity. Will a standard host strain for alpha
complementation, such as DH5 alpha, work for this, or would it be better to
use a strain overproducing the lacZ(delta) M15 deletion? Stratagene sells a
strain,  E. coli SURE, with an F' plasmid with this gene, but they charge
$80 for it - I'd rather not shell out that much if an extract from DH5
alpha would work, or if a similar strain is freely available elsewhere.

Thanks,

John Rebers
Department of Cellular Biology
University of Georgia
Athens, GA  30602
jrebers at nmu.edu
706-542-0802 (voice)
706-542-4271 (FAX)



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