Transformation with ADE2
stahl at infobiogen.fr
Tue Jun 3 18:07:50 EST 1997
> ade2-101 strains?
ade-101 is an ochre mutation !
How is you negative control ? I used to transform trp1-289 (amber), and
the background of reversion is quite high.
Furthermore, there are also potential extragenic mutation which would
supress this allele (tRNAs, and of course omnipotent supressors like
> When I pick these small colonies and streak them out onto fresh plates,
Small colonies after transformation are often rho- clones (but then this
phenotype should not reverse...). Test this on glycerol plates (Anyway,
it'a a good idea to streak your yeast strains from time to time on this
media, to get an homogenous rho+ strain)
Perhaps the explanation of the instability of your [Ade+] pnenotype is
of prionic order : SUP35 may be a prion for yeast, and the prionic fomr,
known as psi is an omnipotent suprressor, but without mendelian
Don't know if this helps or a contrario ;-)
Guillaume Stahl, France
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