synthetic E coli promoter?
chrisb at hgu.mrc.ac.uk
Wed Jun 2 08:16:51 EST 1999
John Dixon (jpcd100 at mole.bio.cam.ac.uk) wrote:
: I wish to place an oligo upstream of the neomycin phosphotransferase
: cassette in a construct of mine to so that it also confers kanamycin
: resistance in E coli. I have a convenient site in between the
: eukaryotic promoter and the neo ATG.
Have you checked your plasmid doesn't already confer kanamycin
resistance in E. coli? Try a range of kanamycin concentrations, say
5--25 ug/ml, with plasmid-free bugs as -ve controls. There may well be
a cryptic bacterial promoter upstream. There ALWAYS is when you DON'T
: What is the mininmum consensus sequence for a promoter active in E coli?
See Duncan's reply.
Chris Boyd | from (but not \ MRC Human Genetics Unit
Christopher.Boyd at hgu.mrc.ac.uk | on behalf of) / Crewe Rd, Edinburgh
http://www.hgu.mrc.ac.uk/Users/Christopher.Boyd EH4 2XU, SCOTLAND
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