Agro strain GV3101 Chloramphenicol resistance

Andrew Bent a-bent at uiuc.edu
Tue Nov 24 14:56:25 EST 1998


Pete -

According to my colleague Stephen Farrand, Biovar 1 agrobacteria mutate to
resistance to chloramphenicol at a very high frequency, and such mutants
are resistant to high levels of chloramphenicol.  You should use
chloramphenicol as a marker in Agrobacterium only with great caution.

Parenthetically, these agro strains also mutate to tetracycline resistance
at a frequency of 10-6 to 10-5, and tetracycline resistance on
RP4/RK2-based plasmids is often poorly expressed in agro.  When using Tc
selection, use only 2.5 ug/ml and pick the small colonies.  Or better yet,
since CaMV 35S promoter drives NPTII effectively in agro, if your plasmid
has that selectable marker system for selection of transformed plants then
it (Km, 25 ug/ml) can be used to select for the plasmid in Agrobacterium.

- Andrew Bent



>Does anyone have any idea what concentration of chloramphenicol works for
>selection with Agro strain GV3101?  The goal is to select for a Cm
>resistant vector in the bacteria.  The closest reference I could find
>indicated 25 ug/ml, but GV3101 (pMP90) grew fine on that.
>
>Many thanks,
>
>Pete
>
>Pete Yorgey
>Dept. of Molecular Biology
>Wellman 10
>Massachusetts General Hospital
>Boston, MA  02114
>
>Phone:  (617) 726-5950
>Fax:    (617) 726-5949



---------------------------------------
Andrew Bent			Phone: 217-244-6308
Department of Crop Sciences	Fax: 217-333-4777
University of Illinois		Office: 389 Madigan Laboratory
1201 W. Gregory Drive		E-mail: a-bent at uiuc.edu
Urbana, IL  61801
http://w3.aces.uiuc.edu/CropSci/faculty/b/bent/





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