Difference between revisions of "Team:SZU-China/Improve"

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Last year,SZU-China submitted part C125-TupA(
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Last year, SZU-China submitted part C125-TupA(
 
<a style="color: #469789;" href="http://parts.igem.org/Part:BBa_K2232003">BBa_K2232003</a>)
 
<a style="color: #469789;" href="http://parts.igem.org/Part:BBa_K2232003">BBa_K2232003</a>)
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, which is the coding sequence (CDS) of a cytoplasmic protein TupA (GenBank: BAB07375.1).This protein catalyzes the conversion of glucuronic acid and L-glutamic acid to polyglucuronic acid and poly-γ-L-glutamic acid.Besides this gene plays a key role in pH homeostasis and increases the alkali resistance of Bacteria.
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, which is the coding sequence (CDS) of a cytoplasmic protein TupA (GenBank: BAB07375.1). This protein catalyzes the conversion of glucuronic acid and L-glutamic acid to polyglucuronic acid and poly-γ-L-glutamic acid.Besides this gene plays a key role in pH homeostasis and increases the alkali resistance of Bacteria.
 
 
 
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This year we decided to add a His-tag at the C-terminus of the protein,so that we can gain this cytoplasmic protein by Histidine-labeled affinity column chromatography.,and analysis at the protein level is easier.
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This year we decided to add a His-tag at the C-terminus of the protein so that we can gain this cytoplasmic protein by Histidine-labeled affinity column chromatography, and analysis at the protein level is easier.
  
 
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We performed Histidine-labeled affinity column chromatograpghy to isolate fusion protein,then verified by SDS-PAGE and Coomassie blue staining.(Fig.1)
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We performed Histidine-labeled affinity column chromatography to isolate fusion protein, then verified by SDS-PAGE and Coomassie blue staining. (Fig.1)
  
 
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Revision as of 11:43, 16 October 2018

Improve

Last year, SZU-China submitted part C125-TupA( BBa_K2232003) , which is the coding sequence (CDS) of a cytoplasmic protein TupA (GenBank: BAB07375.1). This protein catalyzes the conversion of glucuronic acid and L-glutamic acid to polyglucuronic acid and poly-γ-L-glutamic acid.Besides this gene plays a key role in pH homeostasis and increases the alkali resistance of Bacteria.

But it’s a pity that there was no data at the protein level in 2017 SZU-China iGEM’s project.

This year we decided to add a His-tag at the C-terminus of the protein so that we can gain this cytoplasmic protein by Histidine-labeled affinity column chromatography, and analysis at the protein level is easier.

We performed Histidine-labeled affinity column chromatography to isolate fusion protein, then verified by SDS-PAGE and Coomassie blue staining. (Fig.1)