I meet a puzzle in R gene identifing

Xiang Shi shixiang at pubms.pku.edu.cn
Wed Nov 15 10:33:39 EST 2000

I am a graduate student in Peking University, P.R.China. It is my first 
time to be here. I find it a quite good place for me as I can learn and 
discuss some problems with you all. Wish you all good luck with your 
research work.
My work focused on cloning R gene from Rice based on homology sequence of 
Nucleotide Binding Site (NBS). Using degenerate oligonucleotide primers 
designed by the conserved regions of NBS domain of known R-genes and the 
polymerase chain reaction (PCR), I have amplified several rice NBS 
sequences. Furthermore I acquire a novel rice R-gene analog, which contains 
a complete open reading frame (ORF) by screening the rice BAC library. The 
deduced amino acid sequences of the analog demonstrated significant 
sequence similarity with known R-gene sequences lodged in public databases.
The puzzle is when I got this novel rice R-gene, I find it is very hard for 
me do further work. I have done southern blot analysis finding there is 
only one copy of this gene is rice. I try to map this gene in rice genome 
but failed. At the same I have done northern blot analysis finding the gene 
has no constitutive expression in rice.
I am not sure what can I do next step to find the possible function of the 
gene. I am now thinking performing expression studies of the gene in rice 
under diverse pressure of disease. At the same time, I am thinking 
transgenic plant approach. But I am not confident that the experiments will 
work, as there are too many NBS sequence in rice.
That is the puzzle I am encountering. Hope someone can give me some 
suggestion or guidance. I really appreciate your assistance.
One more thing, I am a fresh man in research work, I do know papers is a 
good guide in pushing research work going straight ahead, But when I face 
thousands of paper when I find paper in "plant resistance gene" as a key 
word, I am really confused. What should I do, hope you assist you.


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