Transformation with ADE2

Andrei Popov andrei.popov at bbsrc.ac.uk-x
Thu Jun 5 06:14:34 EST 1997


Stephen Cronin wrote:
> 
> Does anyone have experience with integrating vectors using ADE2 as
> a selectable marker for transforming ade2-101 strains?
> 
>    When I transform I get a small number of large white colonies plus
> about ten times as many very small colonies that never reach full size.
> When I pick these small colonies and streak them out onto fresh plates,
> I get a mix of large and small colonies.  The vectors I am using are
> based on pASZ11 and pASZ10 (Stotz and Linder, (1990)GENE 95:91-98). I
> have been using a Lithium acetate transformation protocol and selecting
> on minimal media with the appropriate supplements.  Have you seen this
> transformation phenotype before, or something similar?
> 
> Thanks,
> Stephen Cronin
> scronin at jeeves.ucsd.edu


I use the same genes and aften get the same. As I also observed
these small colonies (obviously not red) after transformation
with Lys2 marker, I assumed that they represent transient
expression of the auxotrophic gene....
However I remember that once I tried to streak these small colonies
and they did not grow at all on secondary selection plates


with best wishes

Andrei

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