User:Nkuldell/SAGA swap pt 2

Reagents

BBa_Y00073 [[1]]

had DNA 2.0 codon-optimize S. pombe gene SPCC126.04c [[2]] including biobrick ends for expression of ORF in S. cerevisiae This subunit is homolog of S. cerevisiae SGF73 [[3]] subunit of SAGA and deletion of this subunit in S. cerevisiae results in viable cells but no induction of PAU genes under hypoxic conditions.

alignment of S. cerevisiae Sgf73p with S. pombe SPCC126.04c shows alignments btw S.cerevisiae aa 68 to aa 398 with S. pombe seq aa 74 to 318. Sgf73 has C-term region without homology to smaller S.pombe protein. File:Sc Sp Sgf73.pdfand File:Sp Sc Sgf73pseq.pdf
Quite different is the comparison of S. cerevisiae Sgf29 to S. pombe SPBC1921.07c which are proteins of approx the same length (259aa for S.c. vs 244aa for S.p) but which align only at the C-term half. File:Sp Sc Sgf29p .pdf

Codon Usage Comparison using GeneArt tool called Graphical Codon Usage Analyzer [4]

MH176

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
Natalie,
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.
Mark