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TOL plasmids

Paul N Hengen pnh at cockleberry.ncifcrf.gov
Tue Dec 12 15:10:08 EST 1995


Mikhail Alexeyev (malexeyev at biost1.thi.tmc.edu) wrote:

| 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).

: I am not a TOL plasmid expert, but those who work with symbiotic plasmids
: in nitrogen-fixing bacteria (Rhizobium, etc.). for separation of large
: plasmids often use different modifications of the protocol initially
: described by Eckerd (1978-1979). This protocol involves in-gel lysis of
: cells and separation in an agarose gel. AFAIR, the procedure works for
: plasmids up to 500 kb. There are other procedures like one described Kado
: and Liu that involves actual isolation of the plasmid. I beleive, most of
: the protocols and necessary references you can find in Hardy (ed.)
: Plasmids. There were several editions of this book. Sorry, can't provide
: more detailed references.

@article{Eckhardt1978,
author = "T. Eckhardt",
title = "A rapid method for the identification of plasmid
desoxyribonucleic acid in bacteria",
journal = "Plasmid",
volume = "1",
number = "4",
pages = "584-588",
year = "1978"}

@article{Kado1981,
author = "C. I. Kado
     and S. T. Liu",
title = "Rapid procedure for detection and isolation of
large and small plasmids",
journal = "J. Bacteriol.",
volume = "145",
number = "3",
pages = "1365-1373",
year = "1981"}

@article{Adachi1980,
author = "T. Adachi
     and V. N. Iyer",
title = "A procedure for the isolation and purification of
plasmid DNA from Rhizobium meliloti",
journal = "Anal. Biochem.",
volume = "101",
number = "2",
pages = "271-274",
year = "1980"}

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* Paul N. Hengen, Ph.D.                           /--------------------------/*
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