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.
===========================================================================
