User:Nkuldell/SAGA swap pt 2: Difference between revisions
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Codon Usage Comparison using GeneArt tool called Graphical Codon Usage Analyzer [http://gcua.schoedl.de/sequential.php]. Output show on SAGA swap discussion page [[http://openwetware.org/wiki/User_talk:Nkuldell/SAGA_swap]] | Codon Usage Comparison using GeneArt tool called Graphical Codon Usage Analyzer [http://gcua.schoedl.de/sequential.php]. Output show on SAGA swap discussion page [[http://openwetware.org/wiki/User_talk:Nkuldell/SAGA_swap]] | ||
===MH176=== | ===MH176=hypoxic gene expression reporter strain=== | ||
from Mark Hickman in Fred's lab: <br> | from Mark Hickman in Fred's lab: <br> | ||
<b>MATa ura3D0 his3D200 leu2D0 lys2-128d HAP1 DAN3-HIS3 </b><br>geneology note: = yMH36 DAN3-HIS3 #5 (targeted PAU6, went into DAN3) 05/25/05 | <b>MATa ura3D0 his3D200 leu2D0 lys2-128d HAP1 DAN3-HIS3 </b><br>geneology note: = yMH36 DAN3-HIS3 #5 (targeted PAU6, went into DAN3) 05/25/05 | ||
| Line 20: | Line 20: | ||
We can talk in the afternoon. <br> | We can talk in the afternoon. <br> | ||
Mark | Mark | ||
===FY2475 = sgf73::KanMX K/O strain=== | |||
made by Jenny in Fred's lab: <br> | |||
<b> MAT alpha ura3-52 lys2-173R2 leu2D1 arg4-12 his4-917d sgf73D0::kanMX </b> | |||
===FY2474 = sgf29::KanMX K/O strain=== | |||
also by Jenny? <br> | |||
<b> MAT A ura3-52 lys2-173R2 leu2D1 arg4-12 his4-917d sgf73D0::kanMX </b> | |||
Revision as of 07:30, 17 July 2006
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.
Codon Usage Comparison using GeneArt tool called Graphical Codon Usage Analyzer [4]. Output show on SAGA swap discussion page [[5]]
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
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
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
