Gabor Keresztes wrote:
> Dear netters,
>> My aim is to express two similar proteins (wt and mutant) in
> eukaryotic cells. To this aim I chose the pIRES vector which enables the
> expression of two genes from one plasmid. However, when trying to put
> the second insert into the vector the resulting clones will invariably
> (16 out of 16 prescreened by PCR) be rearranged by the time I grow up
> the bacteria (strain DH5alpha) for minipreps.
> Or should I straigh away forget the idea of using a bicistronic
>> Gabor Keresztes, Ph.D
That' s right: forget it.
yu may even get problems if these two inserts are on different plasmids.
clemens suter-crazzolara, PhD