Cloning Vector Help !

Klaus Salger salger at wap1.zi.biologie.uni-muenchen.de
Mon Aug 7 17:24:52 EST 1995


Richard C. Ogden (bss053 at bangor.ac.uk) wrote:

: HELP !!
:        I am attempting to clone a gene from a Pseudomonad bacterial 
: strain. Other anologues of the gene have been studied and it is reported 
: to suffer deletion events and possible toxicity problems in high copy 
: number vectors, such as pUC. They seem reasonably happy in pBR322 
: derivatives or low copy number vectors such pKT230.
:        The problem is pKT230 is not very nice to work with and I need a 
: SmaI site in the vector, ruling out pBR322. Could anyone recommend a 
: possible vector and a supplier ? Any help would be much appreciated !

Richard,
you could try pT7-7. It has a Col EI ori (low copy) and a SmaI site in the
MCS. It's also rather small - 2,5kb. We recently got it from Amersham/USB.
Have a look into the 1995 USB catalog for map and list of RE sites.

Another alternative would be pACYC. It is much bigger (4kb) and carries a
p15A ori. It has two resistance markers Amp and Kan. The SmaI site lies in
the middle of the Kan cassette. It's available from NEB for the cost of
shipping (see NEB catalog).

Hope this helps
  Klaus

--
Klaus Salger                phone : +49 (0)89 5902 -502
Zoologisches Institut       FAX   :                -450
AG MacWilliams              e-mail: salger at zi.biologie.uni-muenchen.de
Luisenstr. 14
80333 Muenchen
Germany



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