User:Nkuldell/SAGA swap pt 2
had DNA 2.0 codon-optimize S. pombe gene SPCC126.04c [] including biobrick ends for expression of ORF in S. cerevisiae This subunit is homolog of S. cerevisiae SGF73 [] subunit of SAGA and deletion of this subunit in S. cerevisiae results in viable cells but no induction of PAU genes under hypoxic conditions.
MH176=hypoxic gene expression reporter strain
from Mark Hickman in Fred's lab:
MATa ura3D0 his3D200 leu2D0 lys2-128d HAP1 DAN3-HIS3
geneology note: = yMH36 DAN3-HIS3 #5 (targeted PAU6, went into DAN3) 05/25/05
related email note 07.12.06 from Mark
I only have PAU5-URA3 or DAN3-HIS3. The DAN3 reporter is tighter on plates, perhaps because the His phenotype is tighter in general. I have not checked whether SGF73 is required for PAU5-URA3 or DAN3-HIS3 induction specifically. But I do know that in an sgf73 delta mutant, there is no hypoxic PAU expression. So I infer that PAU5 and DAN3 expression is affected. We can talk in the afternoon.
FY2475 = sgf73::KanMX K/O strain
made by Jenny in Fred's lab:
MAT alpha ura3-52 lys2-173R2 leu2D1 arg4-12 his4-917d sgf73D0::KanMX
FY2474 = sgf29::KanMX K/O strain
also by Jenny?
MAT A ura3-52 lys2-173R2 leu2D1 arg4-12 his4-917d sgf73D0::KanMX
Phenotype MH176 (~wt) v FY2475 (sgf73::KanMX) v FY2474 (sgf29::KanMX)
First spot where individual colonies seen is listed
Better wt control would be FY3 since FY strains are all hap1
|Plate type||Temp||MH176 (~wt)||FY2475 (sgf73::KanMX)||FY2474 (sgf29::KanMX||comments|
|YPD||30° 3d||6||6||6||Mark has seen 73 KO grow more slowly doing growth curve|
|YPgal||30° 3d||6||4 (repeat this?)||6|