shuttle of plasmids

Magda Smoor
Tue Feb 20 05:36:11 EST 1996

Can anybody comment on the following:

I'm using the yeast two hybrid system of Clontech (pAS2 and pGAD10 library)
and to analyse blue colonies I shuttle the plasmids to E. coli (XL1 
blue). It works fine and I never have problems in getting the library 
plasmid. However, I noticed that the bait vector (pAS2 construct) is only 
present in very low number of E. coli colonies. (<< 1% of the colonies 
contain the bait vector.) Does anybody have an explanation for this? 
Could it be yeast or E. coli strain dependent? I've seen this using 
different constructs.

Magda Smoor
Dept of Human Genetics
Leiden University

mail: magda at ruly46.MedFac.LeidenUniv.NL

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