[Arabidopsis] AtCBF2 salk line mutant
(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?
Discover Live.ca, the search engine by the creators of Windows Live
More information about the Arab-gen