[Arabidopsis] question on over-expression

[Y-Q Jiang]

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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