[Arabidopsis] question on over-expression
jiangyqcn at hotmail.com
Sun Apr 9 16:18:19 EST 2006
When overexpressing one gene under CaMV35S promoter in pCAMBIA vector,
should clone the cDNA with 5UTR 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
5UTR, coding region and 3UTR), 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.
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