Transfer of genes to E.coli chromosome

Dr. Duncan Clark Duncan at genesys.demon.co.uk
Mon Jun 28 15:57:36 EST 1993


Hi Netters

Does anyone have a good reliable method for moving a gene onto the E.coli
chromosome and keeping it there. I want to put a single copy of a trp promotor
controlled lambda cI repressor gene onto the chromosome. (it allows induction
of PR or PL promotors by addition of tryptophan to the media, by switching off
the CI gene. One can therefore induce at low temperatures and not have to heat
shock.) I have the gene on a colE1 based plasmid but don't want to stick the
replicon or it's resistance gene on the chromosome. 

Alternately I have the gene on an F' but the F' is selectable by lac but for
what I want to do I have to use a lac+ E.coli and therefore lose the selection. 

Many thanks

Duncan

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 Duncan Clark                       | Internet:   duncan at genesys@demon.co.uk
 GeneSys Ltd.                       | Compuserve: 100015.1406 at compuserve.com
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