pCDNAI and pCDM8 problems in stable transfection ?
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
> 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.
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).
Ohio State Neurobiotechnology
beall.3 at osu.edu
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