Looking for a vector to make tnx fusions to lacZ in E. coli

Bernard P. Murray, PhD bpmurray*STUFFER* at socrates.ucsf.edu
Thu Oct 8 20:59:45 EST 1998


In article <6vj933$q4i$1 at nntp.ucs.ubc.ca>, rewhlab at cmdr.ubc.ca (rewhlab) wrote:

> Hi,
> 
> Does anyone have a small (less than 8 kb) plasmid that I can use for making 
> transcriptional fusions to lacZ.  THe experiments I want to do are to look at 
> the transcriptional activity of several gene promoters in an E. coli 
> background.
> 
> If you have such a vector and would be willing to send me the strain or
vector 
> please reply to rewhlab at cmdr.ubc.ca

Am I missing your intent or can you not simply use pGEM or
pBluescript and swap out the Lac promoter and replace it
with your promoter of interest?  I realise that you'll
only be expressing the alpha peptide under your control
but this should give you the activity you need in
a delta(lacZ)M15 host.  You may have to fiddle a bit
to introduce a MCS equivalent upstream of the LacZ
start codon but the vectors should still stay at about 3 kb.
     Sorry if I misunderstood,
          Bernard
-- 
Bernard P. Murray, PhD
Dept. Cell. Mol. Pharmacol., UCSF, San Francisco, USA



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