Prokaryotic expression, purification and activity analysis of recombinant human serine protease inhibitor Hespintor Kazal Domain / 生物工程学报
Chinese Journal of Biotechnology
; (12): 1607-1616, 2013.
Article
在 Zh
| WPRIM
| ID: wpr-242432
Responsible library:
WPRO
ABSTRACT
Hespintor is an unknown function protein that was got from hepatoblastoma cell lines HepG2 by suppression subtractive hybridization technique (SSH), sequence analysis showed that the protein is a new member of secretory type of Kazal type serine protease inhibitor (Serpin) family, and has high homology with esophageal cancer related gene 2 (ECRG2). The coding sequence of Hespintor's Kazal domain was subcloned into prokaryotic expression vector pET-40b(+), then transformed into Rosetta (DE3). A recombinant protein about 42 kDa in the form of inclusion body was optimization expressed by inducing with 0.25 mmol/L IPTG, 30 degrees C for 5 h. and its specificity was confirmed via Western blotting. The recombinant protein was purified by metal chelate affinity chromatography (MCAC) and anion-exchange chromatography. The preliminary experimental result showed that the recombinant protein can inhibit trysin hydrolysis activity specifically. The result clearly demonstrated that Hespintor, as a novel member of Serpin, would be valuable in developing anti-tumor agents.
全文:
1
索引:
WPRIM
主要主题:
Recombinant Proteins
/
Serine Proteinase Inhibitors
/
Classification
/
Escherichia coli
/
Proteinase Inhibitory Proteins, Secretory
/
Hep G2 Cells
/
Serine Peptidase Inhibitors, Kazal Type
/
Genetic Vectors
/
Genetics
/
Metabolism
限制:
Humans
语言:
Zh
期刊:
Chinese Journal of Biotechnology
年:
2013
类型:
Article