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WAT11 酵母菌株 BioVector NTCC质粒载体菌种细胞基因保藏中心

  • 价  格:¥19862
  • 货  号:WAT11 酵母菌株
  • 产  地:北京
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WAT11 酵母菌株 BioVector NTCC质粒载体菌种细胞基因保藏中心
The ATR1 and ATR2-1 open reading frames were isolated from pUC19/ATR1 and pUC19/ATR2-1
as BamHI-EcoRI andBamHI-BamHI fragment, respectively. TheATR1fragment was inserted at
theBamHI-EcoRI sites of pYeDP110, yielding pATR1/DP110. Similarly, the ATR2-1 fragment was inserted
at the uniqueBamHI site of pYeDP110, and a clone with the proper orientation was selected and named
pATR2-1/DP110. The NotI fragment of each plasmid encompassing URA3 and the ATR coding sequence
was isolated and used to transform WRΔ cells. Since in WRΔ the CPR1 locus is disrupted with
theTRP1 gene, integration events result in a phenotype shift from Ura− Trp+ to Ura+Trp−. This procedure
yielded two new yeast strains, WAT11 (for ATR1) and WAT21 (for ATR2-1). They express
theArabidopsis CPR instead of the yeast enzyme when grown on galactose. On glucose, WAT cells
express no CPR activity.
Analysis of the coupling between any ATR and a plant P450 requires coexpression in yeast. Genomic
integration of the ATR1 and ATR2-1 expression cassettes at the CPR1 locus was considered. For this
purpose, ATR1 and ATR2-1 coding sequences were cloned into pYeDP110 integration vector, yielding
pATR1/V110 and pATR2-1/V110. The pYeDP110 vector places the heterologous coding sequence under
the transcriptional control of GAL10-CYC1 promoter andCPR1 terminator and provides sequences from
both flanking regions of the CPR1 gene that direct the integration at this locus, resulting in the deletion of
most of the CPR1 gene. The ATR1 and ATR2-1 coding sequences were integrated at theCPR1 locus,
resulting in two new yeast strains, WAT11 and WAT21, respectively.
WAT11 genotype
MATα (leu2-3,112 trp1-1 can1-100 ura3-1 ade2-1 his3-11,15)

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