Expression and characterization of soluble recombinant Ulp1p with glutathione S-transferase tag in Escherichia coli.
- Author:
Junhua FU
1
;
Qi WANG
;
Jiechao YIN
;
Mingyao LIU
;
Ning LI
;
Wenbin YAO
;
Guiping REN
;
Lu LI
;
Deshan LI
Author Information
1. Biopharmaceutical Laboratory, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
- Publication Type:Journal Article
- MeSH:
Cloning, Molecular;
Cysteine Endopeptidases;
biosynthesis;
genetics;
Escherichia coli;
genetics;
metabolism;
Fungal Proteins;
biosynthesis;
genetics;
Glutathione Transferase;
biosynthesis;
genetics;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
Saccharomyces cerevisiae;
enzymology;
Solubility
- From:
Chinese Journal of Biotechnology
2010;26(6):837-842
- CountryChina
- Language:Chinese
-
Abstract:
The aim of the study is to obtain an efficient expression of recombinant ubiquitin-like specific protease 1 (Ulp1) by gene engineering. We cloned the Ulp1p, active fragment (403 aa-621 aa) of Ulp1, from Saccharomyces cerevisia, and subcloned into pGEX/Rosetta (DE3) to form an expression plasmid, pGEX-Ulp1p-His6. In order to enhance the solubility of GST-Ulp1p-His6, we purified the fusion protein GST-Ulp1p-His6 by either glutathione S-transferase agarose or Ni-NTA resin chromatography, the purity was up to 98%. We utilized the protein to cleave the SUMO fusions, and the specific activity of GST-Ulp1p-His6 was 1.375 x 10(4) U/mg. This study showed that the recombinant protein GST-Ulp1p-His6 displayed high specificity and activity.
