Part:BBa_K2665011
TDH3-Mangrin-6xHis-CYC
This part coded the mangrin functional domain(71 amino acids). This part also contains TDH3 promoter, Histag, and CYC1 terminator in order to express constantly in yeasts. We used this part for improving halotorelance of yeast.
Usage and Biology
Mangrin is derived from mangrove and it is a shaperon like protein and enhances salt tolerance of Escherichia coli, Yeast, and Tobacco Cells. This part contains only the functional domain(71 amino acids). TDH3 promoter is a constitutive promoter, so this part can express mangrin functional domain constitutively under appropriate condition.
Characterization
Pictures shown above is colonies of S. cerevisiae ΔENA1- strain on SD midium containing 400mM NaCl. In this spot assay, part BBa_K2665011 cloned to a yeast low-copy vector was used.
This result shows that mangrin 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 mangrin also contributes to accumulation of K+ and Na+ in a yeast cell.
In addition, mangrin on the high copy plasmid inhibit growth of yeasts too extensively to form colonies.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 842
Illegal BamHI site found at 936 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Reference
[1] A. Yamada, T. Saitoh, T. Mimura et al. (2002) Expression of Mangrove Allene Oxide Cyclase Enhances Salt Tolerance in Escherichia coli, Yeast, and Tobacco Cells, Plant and cell physiology 903-910
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