Team:SZU-China/Improve
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)