Difference between revisions of "Part:BBa K2765026:Experience"
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We cloned yno1 from the genome of CEN.PK2-1C,then we ligated the gene yno1 with pESC plasmid,and cloned Pgal1-yno1-cyc1t from pESC plasmid.as shown in Fig.2.
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We cloned <i>yno1</i> from the genome of CEN.PK2-1C,then we ligated the gene <i>yno1</i> with pESC plasmid,and cloned Pgal1-<i>yno1</i>-cyc1t from pESC plasmid.as shown in Fig.2.
  
 
[[Image: T--BIT-China--iGEM2018-Partsregulator-Fig.2-targetgene.png |center|400px|]]
 
[[Image: T--BIT-China--iGEM2018-Partsregulator-Fig.2-targetgene.png |center|400px|]]

Revision as of 21:25, 17 October 2018


We cloned yno1 from the genome of CEN.PK2-1C,then we ligated the gene yno1 with pESC plasmid,and cloned Pgal1-yno1-cyc1t from pESC plasmid.as shown in Fig.2.

T--BIT-China--iGEM2018-Partsregulator-Fig.2-targetgene.png

After sequencing verification, we transferred the correct plasmid into yeast for expression. Then we used the galactose induction medium to induce the Gal1 promoter to overexpress the target gene, which increased the ROS content in the Saccharomyces cerevisiae cell.We used a suitable concentration of galactose to induce overexpression of the target gene, resulting in an increase in ROS accumulation compared to the control group.as shown in Fig.3.

T--BIT-China--iGEM2018-Partsregulator-Fig.3-experimentresult.png

References:

[1]Mark Rinnerthalera, Yno1p/Aim14p, a NADPH-oxidase ortholog, controls extramitochondrial reactive oxygen species generation, apoptosis, and actin cable formation in yeast ，2012

[2]Zhu Kaichuan, Zhang Jianhua, Liu Shide. Advances in yeast transcription factor Gal4 [J]. Chinese Journal of Bioengineering, 2011 1 1 (01): 81-85.


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