Optimal cDNA length in expression vectors?

Wolfgang Schechinger wolfsc at ibms.sinica.edu.tw
Sat Dec 8 03:45:36 EST 2001


Bernhard, 

in my experince with pCDNA3 (and also in general), just using the sequence 
from start to stop works fine. If you add 5' UTR, you'll incerease the 
distance to the CMV promoter which is upstream and maybe insert some undesired 
regulative elements. 

Adding a Kozak consensus element around the ATG (eg gccgccaccATGg) could 
enhance translation.

regards, 
Wo


> I have a basic question regarding expression vectors.
> I would like to clone cDNA's into an expression vector (e.g. pcDNA3) in
> order to do transfection experiments. Clearly, in order to get
> functional protein, the inserted cDNA should contain the whole sequence
> from start (ATG) codon to stop codon. But how does the length of any
> included  5'UTR (or 3'UTR) influence optimal transgene expression? When
> I clone a gene of interest into the MCS of a commercial available
> expression vector, is it ok to start directly with ATG?
> Thanks a lot for any comments
> Bernhard
> 
> 


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Dr. Wolfgang Schechinger
Institute of Biomedical Sciences
Academia Sinica, Taipei, Taiwan R.O.C.
e mail wolfsc at ibms dot sinica dot edu dot tw
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