Difference between revisions of "Part:BBa K2765021:Design"
 
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https://static.igem.org/mediawiki/2018/8/88/T--BIT-China--PartRegulator_yno1_Fig2_Overexpress.png
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To determine whether overexpression of <i>yno1</i> can induce the endogenous ROS accumulation in yeast, we constructed an expression plasmid based on pESC-Leu, in which the cloned <i>yno1</i> is driven by GAL1 promoter, and thus the target gene is induced by galactose and repressed by glucose.
  
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Fig.2 overexpress yno1
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[[Image: T--BIT-China--iGEM2018-Partsregulator-4.png |center|300px|]]
  
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https://static.igem.org/mediawiki/2018/a/ae/T--BIT-China--PartRegulator_yno1_Fig3_process_of_Electron_transformation.png
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Obtained gene <i>yno1</i> though clone from yeast genome.
  
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[[Image: T--BIT-China--iGEM2018-Partsregulator-5.png |center|600px|]]
  
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we constructed an expression plasmid based on pESC-Leu, ligase gene <i>yno1</i> into it, We use <i>E.coli</i> as our host cell to construct the
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rcombinant plasmid, then transformed it into <i>Saccharomyces cerevisiae</i>.
  
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Fig. 3 process of Electron transformation
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[[Image: T--BIT-China--iGEM2018-Partsregulator-6.png |center|600px|]]
  
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Overexpression yno1 gene can causes MCA1 dependent apoptosis; likely involved in superoxide-mediated regulation of the actin cytoskeleton; member of a conserved superfamily of NADPH oxidases (NOX enzymes); has similarity to iron/copper reductases (FRE1-8), particularly Fre8p
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Here shows the result of Colony PCR in <i>E.coli</i>.

Latest revision as of 23:43, 17 October 2018

To determine whether overexpression of yno1 can induce the endogenous ROS accumulation in yeast, we constructed an expression plasmid based on pESC-Leu, in which the cloned yno1 is driven by GAL1 promoter, and thus the target gene is induced by galactose and repressed by glucose.

T--BIT-China--iGEM2018-Partsregulator-4.png

Obtained gene yno1 though clone from yeast genome.

T--BIT-China--iGEM2018-Partsregulator-5.png

we constructed an expression plasmid based on pESC-Leu, ligase gene yno1 into it, We use E.coli as our host cell to construct the rcombinant plasmid, then transformed it into Saccharomyces cerevisiae.

T--BIT-China--iGEM2018-Partsregulator-6.png

Here shows the result of Colony PCR in E.coli.