Hi there,

If you want to perform efficient integration then recombination event has to occur either in your gene of interest or in the selection gene. The problem is to discriminate between intégrants (low probability) and plasmid containing cell (high probability) as the selection is the same in both cases. To improve this you could try to transform your episomal vector into an integration vector by deleting the 2µ region.

why not directly use Yeast Integrative Plasmids (YIPs) for expression. there is a huge variety of them. however, be aware that the expression level will be low because the copy number will be 1. 

I agree with the previous two responses. The best solution is probably to move your construct into a related integrating vector and integrate it using the resulting plasmid instead.

have you thought of using a GAP repair technique? you co-transform an (episomal) plasmid linearized and a PCR fragment containing your channel flanked by regions homologous to the cut ends of your plasmids. I have used it in combination with the yeast expressing system described in Geymonat 2007 Gene and got good results expressing cdc5

Integration of an episomal (presumably 2 micron) plasmid into the yeast genome causes instability of the chromosome where the gene has integrated.  So it is not a good idea to integrate an episomal plasmid.  Depending on the structure of your construct, you could potentially convert it to an integrating plasmid by deleting the origin and segregation element of the plasmid you currently are working with, rather than sub cloning the insert.

Malcolm, that's a good point about chromosomal instability.

thanks a lot for the information.