Difference between revisions of "Part:BBa K2765026:Experience"

 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
how you used this part and how it worked out.
 
how you used this part and how it worked out.
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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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https://static.igem.org/mediawiki/2018/d/d8/T--BIT-China--Parts_regulator_Fig.2_target_gene.png
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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.
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https://static.igem.org/mediawiki/2018/4/42/T--BIT-China--Parts_regulator_Fig.3_Experimental_result.png
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References:
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[1]Mark Rinnerthalera, Yno1p/Aim14p, a NADPH-oxidase ortholog, controls extramitochondrial reactive oxygen species generation, apoptosis, and actin cable formation in yeast ，2012
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[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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===Applications of BBa_K2765026===
 
===Applications of BBa_K2765026===

Revision as of 07:22, 17 October 2018


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

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--Parts_regulator_Fig.2_target_gene.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--Parts_regulator_Fig.3_Experimental_result.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.


Applications of BBa_K2765026

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