[Arabidopsis] question on overexpression construct

Y Jiang jiangyqcn at hotmail.com
Fri Sep 1 15:28:35 EST 2006


Dear Arabidopsis researchers,

I recently want to overexpress an Arabidopsis gene in Arabidopsis under 
CaMV35S promoter. I read some references and found most papers amplified and
introduced only the coding region into binary vectors, although a few papers 
used full-length cDNA (contains 5’UTR, coding region(ORF), and 3’UTR). Since 
the gene I am going to overexpress in Arabidopsis contains a short signal 
sequence before the typical ATG start codon(the signal sequence is expected 
to direct the expressed protein to the chloroplast), and I plan to use 
pCABMIA1303 vector and replace the
GUS reporter gene with my gene between NcoI(containing ATG start codon) and 
BstEII, I am wondering it is better to use coding region plus signal 
sequence or the full-length cDNA? Is there any difference between them? Does 
5’UTR of the gene have any influence on the overexpression of the gene?

Thanks for your reply.

Jiang

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