Hi Trent,
Yes I have seen multiple cDNA's rescued from a single two-hybrid positive.
This usually arises from yeast that are inadvertently carried over when you
pick your positives. It can also be related to your prey copy number - but
I think the proquest system is low copy. You can usually get around it by
re-streaking your positives back onto selective media and picking a single
colony from there. pPC86 carries the AMP marker (PNAS 89, pp.5789-5793) - I
expect pPC97 does as well.
Cheers
Martin
Interomex Biopharmaceuticals Inc.
3590 West 41st Ave.
Vancouver, BC
Canada
V6N 3E6
On 6/4/02 12:06 PM, "Trent Bosma" <Trent_member at pathlink.com> wrote:
> Yeast two hybrid (ProQuest system using ResGen Hela cDNA library)
>> 1. The majority of my putative interactors contain more than 1 cDNA. Has
> anyone experienced similar results and found a cure apart from changing
> my bait?
>> 2. Has anyone successfully extracted the plasmids out of the controls
> provided with the kit i.e. what is the antibiotic marker of pPC97?. I was
> expecting it to be kanamycin as with pPC86 but it doesn’t appear to be so.
>> Trent J. Bosma PhD.
> LCI/NIAID/NIH
> Bldg. 10/11N222
> 301 496 7680
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