cloning toxic genes

Martin Hughes Biochem mjgh at mbfs.bio.cam.ac.uk
Wed Jul 7 04:14:45 EST 1993


In article <9307011940.AA05274 at net.bio.net> N490047 at UNIVSCVM.CSD.SCAROLINA.EDU (Tom MacAllister) writes:
>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


-- 
Martin J. G. Hughes
Department of Biochemistry, University of Cambridge, Cambridge, UK.
Email: mjgh at mbfs.bio.cam.ac.uk  OR  hughes at rkna50.riken.go.jp



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