pseudomonas vectors

Mark S. Strom strom at listeria.nwfsc.noaa.gov
Wed Dec 6 16:38:10 EST 1995


Don Chen wrote:

 
> Does anyone have information regarding cloning vectors for Pseudomonas
> spp that can also work in Ecoli?  The type of vectors include, but not
> restricted to, pBR-type cloning vectors, shuttle vectors, vectors with
> transposon cassettes, and transducing phage vectors. 

There are several cloning vectors.  The ones I'm most familiar with (from my P. 
aeruginosa days) include an IncQ vector derived from RSF1010 (actually a series of 
vectors) called pMMB66EH, pMMB66HE, pMMB67EH, and pMMB67HE.  These are good shuttle 
vectors, encode beta-lactam resistance (amp in E. coli, carbenicillin in Pseudomonas), 
have multiple cloning sites, and carry a tac promoter (for high expression in E. coli), 
as well as a constituitive lacIq gene (induction of the tac promoter with IPTG). These 
hae worked really well for high level expression in both E. coli and P. aeruginosa. The 
reference for these is Furste, et. al., 1986. Gene 48:119-131. Several IncP derivatives 
have also been used but I don't have those references handy.  

Several transposons have been used to mutagenize Pseudomonas--again the ones I'm 
familar with were used in P. aeruginosa and were derivatives of Tn5, TnphoA, usually 
put on vectors (ColE1) that could be mobilized and transferred into P. aeruginosa by 
conjugation but wouldn't replicate.  You might want to post this in the 
bionet.organisms.pseudomonas newsgroup where you'll probably get more definitive 
information.

Good luck.

--
Mark S. Strom, Ph.D.                 (206) 860-3379  
Utilization Research Div.            (206) 860-3394 (Fax)
Northwest Fisheries Science Ctr.     mstrom at sci.nwfsc.noaa.gov   
NMFS/NOAA                            strom at listeria.nwfsc.noaa.gov
2725 Montlake Blvd. E.              
Seattle, WA  98112-2097
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