Difference between revisions of "Part:BBa K2665013"

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	==Characterization==		==Characterization==
−	<!--續さん　ここにお願いします-->	+	[[File:T--Kyoto--AtNHXS1.png|400px]] <br>
		+	[[File:T--Kyoto--cont.png|400px]]<br>
		+
		+	Pictures shown above is colonies of<i> S. cerevisiae</i> &Delta;ENA1- strain on SD midium containing 400mM NaCl. In this spot assay, part BBa_K2665013 cloned to a yeast low-copy vector was used. <br>
		+	This result shows that AtNHXS1 greatly contributes to salt tolerance of yeasts.
		+
		+
		+
		+	[[File:T--Kyoto--K_con.jpeg|400px]] <br>
		+	[[File:T--Kyoto--Na con.jpeg|400px]]<br>
		+	The graph shown above is K+ or Na+ concentration in cells of<i> S.cerevisiae</i> &Delta;ENA1&Delta;NHA1 strain. Each gene in the graph was cloned to yeast vectors, which were introduced to yeasts. The word “high” means high copy plasmid and the word “low” means low copy plasmids. These transformed yeasts are cultured in SD containing 400mM NaCl. Detailed data are on our wiki.'''[http://2018.igem.org/Team:Kyoto Kyoto2018]'''<br>
		+	These results show that AtNHXS1 contributes to accumulation of K+ and Na+ in a yeast cell.
		+	In addition, yeast harboring AtNHXS1 in high copy plasmids showed significantly low growth rate
		+
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Revision as of 22:56, 17 October 2018

TDH3-AtNHXS1-6xHis-CYC

AtNHXS1 is a Na+/H+ antiporter which is located in the membrane of vacuoles and that original comes from Arabidopsis thaliana.

We assembled BBa_K2225000 with TDpromoter and CYC1 terminator to express the protein, and we added His-tag in case of Western blotting.

Usage and Biology

This part sequence can be used directly because it contains promoter and terminator, but promoter is TDH3 promoter so it can't express in E. coli. AtNHXS1 is a Na+/H+ antiporter which is located in the membrane of vacuoles, so we used this part in order to increase the amount of Na+ uptake of yeast.

Characterization

400px

Pictures shown above is colonies of S. cerevisiae ΔENA1- strain on SD midium containing 400mM NaCl. In this spot assay, part BBa_K2665013 cloned to a yeast low-copy vector was used.
This result shows that AtNHXS1 greatly contributes to salt tolerance of yeasts.

The graph shown above is K+ or Na+ concentration in cells of S.cerevisiae ΔENA1ΔNHA1 strain. Each gene in the graph was cloned to yeast vectors, which were introduced to yeasts. The word “high” means high copy plasmid and the word “low” means low copy plasmids. These transformed yeasts are cultured in SD containing 400mM NaCl. Detailed data are on our wiki.[http://2018.igem.org/Team:Kyoto Kyoto2018]
These results show that AtNHXS1 contributes to accumulation of K+ and Na+ in a yeast cell. In addition, yeast harboring AtNHXS1 in high copy plasmids showed significantly low growth rate

Sequence and Features

Reference

BBa_K2225000

Sequence and Features