cloning toxic genes

Tom MacAllister N490047 at UNIVSCVM.CSD.SCAROLINA.EDU
Thu Jul 1 14:33:44 EST 1993


We are working with a gene from Pseudomonas that has a high G/C content and
this along with the activity of the encoded protein seems to make it toxic to
E. coli.  We are using the lambda Zap cloning vector with antibody detection.
We have had good luck in recovering primary positives, but many do not enrich.
Those that do enrich give both low yields and multiple unrecognizeable
products.  We have had some luck with similar problems in the past, but this
is  particularly bad.  If anyone has any experience with this sort of thing or
simply a good idea with no basis in reality, we would love to hear it.

Thanks,

Tom MacAllister  <n490047 at UNIVSCVM.CSD.SCAROLINA.EDU>
Natarajan Sethuraman



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