[Arabidopsis] question on over-expression

Y-Q Jiang jiangyqcn at hotmail.com
Sun Apr 9 16:18:19 EST 2006


Hello,

When overexpressing one gene under CaMV35S promoter in pCAMBIA vector, 
should clone the cDNA with 5’UTR or is it ok with the coding region(i.e. 
from ATG to the stop codon)? I know it is ok with the whole cDNA(including 
5’UTR, coding region and 3’UTR), but I am not sure if it is also ok with 
only the coding region? The vector has Nos-ter. I once constructed only the 
coding region with one tag under CaMV35S, and did not detect the expression 
of the protein with anti-tag antibody, although RT-PCR showed that gene was 
transcribed. Should I clone the whole cDNA when going to overexpress it?

Your reply is appreciated.

Thanks.

Jiang

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