Cm-R or Gm-R cassette?
brad at corona.med.utah.edu
Fri Aug 18 15:52:59 EST 1995
In article <t.barnett-180895155956 at mg1_44.med.utas.edu.au>,
t.barnett at path.utas.edu.au () wrote:
>I am after a chloramphenicol-R or gentamicin-R cassette
There are a couple of things that you can do. If you are in a hurry, you
can get a nice cat gene from pACYC184. The gene is contained on a 1325 bp
fragment from a Hae II digest, it's the largest band, you can then blunt
with knenow and clone in to a blunted site of your choice. It sounds
complicated, but it's not that bad because all your non-recombinants die.
The down side is that there are no additional transcriptional terminators
after the stop and you could possibly have some sort of promoter activity.
I don't know about expression in Aeromonas spp., but the original cat gene
in pACYC184 came from Tn9.
If you have some time, there is a recent paper out by Alexeyev, Shokolenko
and Croughan. 1995. Improved antibiotic-resistance gene cassettes adn omega
elements for Esscherichia coli vector construction and in vitro
deletion/insertion mutagenesis. Gene, 160:63-67. They describe some nice
omega elements with cam and gent resistance genes. These constructs come
is several flavors, with a plethora of available cloning sites.
Correspondence is directed to:
Dr. T. P. Croughan
Rice Research Station
P.O. Box 1429
Crowley, LA 70527-1429
e-mail:agexp86 at lsuvm.sncc.lsu.edu
(I first read about these vectors on this newsgroup, I can't remember who
posted it, but they should get an attaboy also.)
Brad Nicholson |"If it worked the first time, it wouldn't be
Department of Pathology | research."...Brad Nicholson
University of Utah | Live from behind the Zion Curtian.
Salt Lake City, UT 84132 |
brad at corona.med.utah.edu |
or: (801)-581-4365 | My opinions are solely my own.
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