title: [Expression and characterization of soluble recombinant Ulp1p with glutathione S-transferase tag in Escherichia coli].
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.
authors.affiliation: Biopharmaceutical Laboratory, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Junhua
authors.initials: J
authors.lastname: Fu
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.forename: Jiechao
authors.initials: J
authors.lastname: Yin
authors.forename: Mingyao
authors.initials: M
authors.lastname: Liu
authors.forename: Ning
authors.initials: N
authors.lastname: Li
authors.forename: Wenbin
authors.initials: W
authors.lastname: Yao
authors.forename: Guiping
authors.initials: G
authors.lastname: Ren
authors.forename: Lu
authors.initials: L
authors.lastname: Li
authors.forename: Deshan
authors.initials: D
authors.lastname: Li
mesh_terms: D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000235:genetics; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012995:Solubility
chemical_list: D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D003546:Cysteine Endopeptidases; C118083:Ulp1 protease