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Biochemistry (Moscow)

, Volume 75, Issue 4, pp 481–485 | Cite as

Production of bioactive human hemangiopoietin in Escherichia coli

  • Jie Zhang
  • Jian-Feng Li
  • Shuang-Quan ZhangEmail author
Article
  • 40 Downloads

Abstract

To devise an efficient approach for production of human hemangiopoietin (hHAPO), the gene of hHAPO was synthesized and subcloned into the pSUMO vector with a SUMO tag at the N-terminus. The expression construct was then transformed into the expression strain E. coli BL21(DE3). The fusion protein was expressed in soluble form and identified by SDS-PAGE and Western blotting. The fusion protein was purified to 90% purity by metal chelate chromatography with a yield of 45 mg per liter fermentation culture. The SUMO tag was removed by cleavage with SUMO protease at room temperature for 1 h, and the hHAPO was then re-purified by the metal chelate chromatography. Finally, about 21 mg hHAPO was obtained from 1 liter of fermentation culture with no less than 95% purity. The recombinant hHAPO significantly stimulated the proliferation of human umbilical vein endothelial cells.

Key words

E. coli fermentation hemangiopoietin Ni-NTA purification SUMO 

Abbreviations

(h)HAPO

(human) hemangiopoietin

HUVEC

human umbilical vein endothelial cells

IPTG

isopropyl-β-D-1-thiogalactopyranoside

SUMO

small ubiquitin-related modifier

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Copyright information

© Pleiades Publishing, Ltd. 2010

Authors and Affiliations

  1. 1.Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Science CollegeNanjing Normal UniversityNanjingJiangsu, P. R. China

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