Gentamycine resistance

Brad Nicholson brad at corona.med.utah.edu
Tue Jul 11 15:03:43 EST 1995


To: vachon at bio.grenet.fr (Gilles VACHON)
From: brad at corona.med.utah.edu (Brad Nicholson)
Organization: Department of Pathology
Subject: Gentamycine resistance

You wrote:

> Hi all,
> 
> I am trying to transform E. coli with a vector carrying a Gentamycine
> resistance gene.
>snip<
 
Hello Gilles,
I have used Gentamycin at 10 ug/ml successfully.  In my hands, the cosmid 
pAM2 (a derivative of pLAFR1 with the Gent cassette from pLB41 cloned in) 
worked ok on LB, although it was a better Kan resistance marker than Gent. 
I believe that the gene is an 3-N-Acetyl-transferase, AAC(3). Ref: Foster, 
T. J.. 1983. Microbiological Reviews. Plasmid Determined Resistance to 
Antimicrobial Drugs and Toxic Metal Ions in Bacteria. 47:361-409.
Which vector are you using and do you know which Gentamycin resistance 
cassette is present?  If the copy # of your construct is very low then you 
may have to drop the amount of Gent present.

Good Luck,
Brad

===========================================================================
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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