Stable COS cell line

eggermont at molbiol.ox.ac.uk eggermont at molbiol.ox.ac.uk
Wed Jul 28 11:04:59 EST 1993


Hello,

I am planning to make a stable COS cell line using G418 selection, but I am 
not sure whether the vector containing the Neo resistance unit may have SV40 
ori sequences. I was once told that excessive replication due to large 
TAg/SV40 ori interactions lead to cell death due to plasmid overload. clearly, 
this would jeopardise the generation of stable clones. To overcome this I 
could use a neo vector without SV40 sequences or ,alternatively, I can 
inactivate the SV40 ori by SfiI digest.
So, has anyone tried and succeeded making stable COS cells? If so, did you 
bother about SV40 ori sequences in the vector? 
Any hints, tips or suggestions are very welcome. Thanks,

Jan Eggermont



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