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TOL plasmids

Kevin O'Donnell odonnell at sasa.gov.uk
Wed Dec 13 12:22:28 EST 1995


In article <4ai9os$up at news2.ucsd.edu>, rcaspi at sdcc3.ucsd.edu (Ron 
Caspi) says:
>
>Hi, is there a TOL plasmid expert out there?
>
>I am working with a strain of P. putida, and would like to check for
>the presence of such a plasmid. Does anybody have a protocol that will
>let me visualize such a plasmid on a gel?  (unfortunately, pulse field
>is not available).
>
>Many thanks in advance
>
I did my PhD on a TOL plasmid from Pseudomonas putida. I used 
sucrose gradients to isolate my plasmid preps - got good enough DNA to 
produce good restriction patterns, clone specific fragments etc.

I don't have the exact reference but it will be found in any of the TOL 
papers from the lab I was in - e.g. Keil et al, J.Bact vol. 163:248-255

The plasmids when you grow the bacteria up on nutrient broth prior to the 
sucrose gradient procedure - but growing them on benzoate seems to 
select all kinds of mutant phenotypes!

Good luck.



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