pCDNAI and pCDM8 problems in stable transfection ?

Clifford Beall beall.3 at OSU.EDU
Wed Jul 5 11:53:33 EST 1995


In article <marcvdc.130.2FF9500F at lmb1.rug.ac.be>, marcvdc at lmb1.rug.ac.be wrote:

> Dear Netters,
> 
> 
> It seems that there are problems with the stable transfections of genes 
> cloned in pCDM8.  Although the transient transfections of the gene gave 
> high expression levels, no protein was detected in the stable transfectans.  
> The same was done with an biological inactive form of the protein and this 
> gave the same results. So, it is not because of the toxicity of the gene 
> expressed.
> 
> Now I wonder wether someone had the same problems with pCDM8.  Are the same 
> problems seen in the pCDNAI (in vitrogen)?  Maybe, I will use this vector in 
> the future but then I want to be sure not to encounter the same problems.
> 
> Please respond also to me personal.
> 
> Thanks,
> 
> Marc

Dear Marc,

You need a selectable marker to get stable transformants, because only
about 1 in 10~6 cells stably integrates introduced DNA. pCDM8 does not
contain such a marker.  You can use pcDNA 3, and select with G418 (you can
find details in Current Protocols in Molecular Biology or other source).

Good luck,
Cliff Beall
Research Scientist
Ohio State Neurobiotechnology
beall.3 at osu.edu



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