cos stable transfection

Mark O'Reilly Mark_O'Reilly at sandwich.pfizer.com
Tue Oct 29 17:26:16 EST 1996


Hugo wrote:
> 
> Dose anyone have experience on stable transfection on Cos cells?  What should b
> e careful in doing this?  Is it necessary to keep the expression of foreign gen
> e low?  If yes, it would be difficult to detect its expression.  Then, it would
>  be very difficult to do further studies on the protein.  Any suggestions?


It is not absolutely necessary to remove the SV40 ori from your vectors, 
or linearise the DNA, prior to transfection as stable cell-lines have 
been successfully isolated from SV40 LTAg expressing cells when 
transfected with SV40 ori-containing vectors. The plasmid has been 
stabily maintained at ~750copies per cell (Gerard & Gluzman, 1985 MCB 
5,11 3231).



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