Methotrexate selection of transgenics

Gregory Armstrong armstrong at aeolus.ethz.ch
Fri Jul 15 11:30:54 EST 1994


Dear Netters,

A colleague of mine with no access to the network has some questions about
methotrexate as a selectable marker for transgenic Arabidopsis.  Seeds from
his primary transformants yield green, presumably resistant seedlings that 
do not develop beyond the cotyledon stage under selective pressure (I don't 
know the methotrexate concentration used).  These seedlings also often grow
along the surface of the agar.  An additional problem is that the germination 
rate under selective pressure is low.  In a control transformation GUS
staining suggests that the seedlings are indeed transformed.
	Any advice or comments?  How many of you work with methotrexate
selection?  Thanks in advance!

***********************************************************************

Greg Armstrong
Institute for Plant Sciences
LFW D31
Swiss Federal Institute of Technology (ETH)
CH-8092 Zurich
Switzerland

Telephone: 41-1-632-3700
Telefax: 41-1-252-0829
Email: Armstrong at aeolus.ethz.ch



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