T7 RNA polymerase phage?

is.bbsrc.ac.uk majones at abdn.ac.uk
Fri Jan 9 07:01:42 EST 1998


Brad Pietz <pietz at oncology.wisc.edu> wrote in article
<pietz-ya02408000R0701981458180001 at news.doit.wisc.edu>...
I hope you find the following iformation helpful, 

coli that appears to be
> quite toxic.  I am able to transform JM109 just fine with my pET vector
> construct but am unable to transform any strain containing T7 RNA
> polymerase (BL21 DE3).  I have heard of people infecting JM109 with
> recombinant phage containing T7 polymerase.  Does anyone know of a source
> for these phage (lambda or M13)?

There is a bacteriophage CE6 (cat. no. 69390)which can  be used to
introduce the T7 polymerase.
This is a lammda phage recombinant. 
Studier and Moffat 1986 J. Mol. Biol. 189,113
This can be used with host HMS174, where T7 polymerase production is so
high 
it interferes with mnormal phage development. 
 
> I have also read about a group (Appl Microbiol Biotechnol 47:241-245.)
> using another plasmid containing a T7 promoter to sop up leaky T7
> polymerase in the cell.  Does anyone know of a source for this plasmid? 

You may want to ,look up studier et al. 1990, Meth. enzymology 185:60-89;
dubenodorff and studier 1991  J. Mol. Biol 219, 45-59. which deals with a
vectors, with T7lac promotors ( the lac operator downstream 
of of the T7 promotor ).  
I have not tried these systems at all, but have got this information from
the novagen ( Novagen US:800 526-7319, distributed by someone else in the
UK.) manual on  pET vectors, so you may want to get hold of that from them
as it has more information and references. 

I hope this helps

Mike Jones
I.A.H. 
Compton
UK

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