In article <4ai9os$up at news2.ucsd.edu>, rcaspi at sdcc3.ucsd.edu (Ron
>>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!