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[Arabidopsis] AtCBF2 salk line mutant

Luoping Yan via arab-gen%40net.bio.net (by yanluoping At hotmail.com)
Thu Jan 4 18:46:43 EST 2007

Hi Arabidopsis researchers,

I recently want to screen out the homozygous cbf2 gene Salk T-DNA 
insertional mutant lines by PCR according to Salk Institute protocol. The 
Salk lines we ordered from ABRC are SALK_009689c   (LOCN 1000-Promotor) and 
Salk_025195(LOCN 300-UTR5). However I failed to get true knock-out mutants 
after running RT-PCR, although PCR results using LBb1/RP and LP+RP showed 
that I got the putative homozygous lines. This can be partly explained by 
the fact that T-DNA insertion in the promoter region may not influence the 
transcription of CBF2 in Salk_009689, but I expect T-DNA insertion near ATG 
start codon in Salk_025195 should block the transcription of CBF2.

One paper in PNAS(2004): CBF2/DREB1C is a negative regulator of CBF1/DREB1B 
CBF3/DREB1A expression and plays a central role in stress tolerance in
Arabidopsis. In this paper, the authors reported a Salk line HM mutant-no 
detectable CBF2 transcript by Northern blotting. I then asked the 
corresponding author a few months ago but no reply at all. So I have no way 
to know which Salk line they used.

Could anyone working with CBF2 mutant reply?



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