Difference between revisions of "Part:BBa K2665012"
 
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It increases glycerol synthesis which work as compatible solute. So, it may raise osmotolerance in yeast.
 
It increases glycerol synthesis which work as compatible solute. So, it may raise osmotolerance in yeast.
  
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==Usage and Biology==
 
==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 <i>E. coli</i>. If need to use the protein in <i>E. coli</i>, please note that you have to change promoter and terminator. ZrGPD1 is the gene that codes a glycerol-3-phosphate dehydrogenase which controls glycerol synthesis.
 
This part sequence can be used directly because it contains promoter and terminator, but promoter is TDH3 promoter so it can't express in <i>E. coli</i>. If need to use the protein in <i>E. coli</i>, please note that you have to change promoter and terminator. ZrGPD1 is the gene that codes a glycerol-3-phosphate dehydrogenase which controls glycerol synthesis.
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==Characterization==
 
==Characterization==
 
<!-- 續さん、ここに書き込んでください、お願いします-->
 
<!-- 續さん、ここに書き込んでください、お願いします-->
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We used this part in order to enhance the solt tolerance of yeast. We assessed the tolerance in our method(please refer to '''[http://http://2018.igem.org/Team:Kyoto/Protocol our Wiki's Special Protocol]''' ).
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We used this part in order to enhance the solt tolerance of yeast.
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We assessed the tolerance in our method(please refer to '''[http://http://2018.igem.org/Team:Kyoto/Protocol our Wiki's Special Protocol]''' ).<br><br>
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[[File:T--Kyoto--ZrGPD1.png|400px]] <br>
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[[File:T--Kyoto--cont.png|400px]]<br>
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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_K2665012 cloned to a yeast high-copy vector was used. <br>
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This result shows that ZrGPD1 contributes to salt tolerance of yeasts.
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[[File:T--Kyoto--K_con.jpeg|400px]] <br>
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[[File:T--Kyoto--Na con.jpeg|400px]]<br>
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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>
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These results show that ZrGPD1 also contributes to accumulation of K+ and Na+ in a yeast cell.
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Result is showed below.
 
Result is showed below.
  

Latest revision as of 22:29, 17 October 2018


TDH3-ZrGPD1-6xHis-CYC

ZrGPD1 is the gene that codes a glycerol-3-phosphate dehydrogenase which controls glycerol synthesis in Zygosaccharomyces rouxii. It increases glycerol synthesis which work as compatible solute. So, it may raise osmotolerance in yeast.

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. If need to use the protein in E. coli, please note that you have to change promoter and terminator. ZrGPD1 is the gene that codes a glycerol-3-phosphate dehydrogenase which controls glycerol synthesis.


Characterization

We used this part in order to enhance the solt tolerance of yeast. We assessed the tolerance in our method(please refer to [http://http://2018.igem.org/Team:Kyoto/Protocol our Wiki's Special Protocol] ).

T--Kyoto--ZrGPD1.png
T--Kyoto--cont.png

Pictures shown above is colonies of S. cerevisiae ΔENA1- strain on SD midium containing 400mM NaCl. In this spot assay, part BBa_K2665012 cloned to a yeast high-copy vector was used.
This result shows that ZrGPD1 contributes to salt tolerance of yeasts.


T--Kyoto--K con.jpeg
T--Kyoto--Na con.jpeg
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 ZrGPD1 also contributes to accumulation of K+ and Na+ in a yeast cell.


Result is showed below.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1923
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1454
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1129
    Illegal BsaI site found at 1889
    Illegal BsaI.rc site found at 1271

Reference

[1] Hou,Lihua Wang,Meng Wang,Cong Wang,Chunling Wang,Haiyong (2013) Analysis of salt-tolerance genes in zygosaccharomyces rouxii, Applied Biochemistry and Biotechnoloogy 1417-1425