Difference between revisions of "Part:BBa K2765021:Design"
 
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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.
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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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https://static.igem.org/mediawiki/2018/7/7b/T--BIT-China--PartRegulator_yno1_Fig4_Size.png
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[[Image: T--BIT-China--iGEM2018-Partsregulator-4.png |center|300px|]]
  
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Fig.4 The size of yno1 (1713bp)
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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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https://static.igem.org/mediawiki/2018/b/b3/T--BIT-China--PartRegulator_yno1_Fig5_Plasmid.png
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[[Image: T--BIT-China--iGEM2018-Partsregulator-6.png |center|600px|]]
  
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Fig.5 plasmid of yno1 expression
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Here shows the result of Colony PCR in <i>E.coli</i>.
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https://static.igem.org/mediawiki/2018/5/5e/T--BIT-China--PartRegulator_yno1_Fig6_ColonyPCR.png
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Fig.6 result of Colony PCR
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1~7: negative results
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8: positive result (yno1 1713bp)
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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.