yeast strain for disr
tod.lorenz at ghawk.com
Tue Jan 31 20:59:00 EST 1995
-=> Quoting Colin Macdiarmid to All <=-
CM> Hi out there
CM> I need some help from experienced gene-disruptors. I have two genes
CM> which I need to interupt, and want to do this by insertion of the URA3
CM> gene. After that I want to be able to use 5-FOA to reselect for a
CM> strain which has excised the URA3 gene (but retained my disrupted
CM> version of the gene). The genes were both from a library made from the
CM> S288C strain, and strains with this background are sensitive to low
CM> concentrations of 5-FOA on plates with proline as sole nitrogen source.
CM> So ideally I want a diploid yeast strain derived from s288C to do my
CM> disruptions. Anyone have an idea which strain to use? Can I get it
CM> from the YGSC? Please post replies or send direct to me.
Is the gene(s) disruption lethal? It should not matter what
the source of the clone bank is, just the background of the yeast
being transformed...try the Berkeley yeast stocks for other ura3
strains to use. Why the need for proline as a nitrogen source
out of curiosity? Why not just excise the URA3 gene out with a
restriction enzyme if the plasmid is mapped and get around all
the FOA problems?
/^\/^^^^\/^\ R. Todd Lorenz
| # @ | Ellicott City, MD
\ :: / T.LORENZ at GENIE.GEIS.COM
___ Blue Wave/QWK v2.12
o Standard disclaimer: Views expressed are strictly those of the author.
o From The GreyHawk BBS - Columbia, MD +1-410-290-3260 +1-410-720-5083
More information about the Yeast