Cloning, Expression and Purification of a novel anti-angiogenic factor-Tumstatin

摘要

blot identified it’s right.Gene Therapy and Molecular Biology Vol 10, page 245Tumor progression may be controlled by various fragments derived from noncollagenous 1 (NC1) C-terminal domains of type IV collagen. Tumstatin peptide is an angiogenesis inhibitor derived from type IV collagen and inhibits in vivo neovascularization induced by vascular endothelial growth factor (VEGF), Here, we firstly showed the expression, cloning and purification of tumstatin from Chinese abortus kidney tissue by RT-PCR, and the construction of pET-His expressive plasmid in prokaryotic cells. Also its’ activity was examined by mouse antiserum against native Tumstatin. The results indicated E.coli BL21(DE3)plysS/ pET-His-tumstatin was induced 3 h by 0.2 mmol/L IPTG at 30°C, and got a high-level expression of 37.9%. The Tumstatin protein was one-step purified by immobilized metal-chelating affinity chromatography (IMAC) and its purity was above 95%. Western