[Arabidopsis] question on overexpression construct
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 5UTR, coding region(ORF), and 3UTR). 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
5UTR of the gene have any influence on the overexpression of the gene?
Thanks for your reply.
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