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Journal of Zhejiang University SCIENCE B 2009 Vol.10 No.2 P.79~86

http://doi.org/10.1631/jzus.B0820385


Expression and purification of bioactive high-purity human midkine in Escherichia coli


Author(s):  Zhong-hui ZHANG, Li-juan DU, Di XIANG, Shun-ying ZHU, Ming-yuan WU, Hui-li LU, Yan YU, Wei HAN

Affiliation(s):  Laboratory of Regenoromics, School of Pharmacy; more

Corresponding email(s):   weihan@sjtu.edu.cn

Key Words:  Expression, Purification, Human midkine, Escherichia coli


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Zhong-hui ZHANG, Li-juan DU, Di XIANG, Shun-ying ZHU, Ming-yuan WU, Hui-li LU, Yan YU, Wei HAN. Expression and purification of bioactive high-purity human midkine in Escherichia coli[J]. Journal of Zhejiang University Science B, 2009, 10(2): 79~86.

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author="Zhong-hui ZHANG, Li-juan DU, Di XIANG, Shun-ying ZHU, Ming-yuan WU, Hui-li LU, Yan YU, Wei HAN",
journal="Journal of Zhejiang University Science B",
volume="10",
number="2",
pages="79~86",
year="2009",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B0820385"
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%T Expression and purification of bioactive high-purity human midkine in Escherichia coli
%A Zhong-hui ZHANG
%A Li-juan DU
%A Di XIANG
%A Shun-ying ZHU
%A Ming-yuan WU
%A Hui-li LU
%A Yan YU
%A Wei HAN
%J Journal of Zhejiang University SCIENCE B
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%P 79~86
%@ 1673-1581
%D 2009
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B0820385

TY - JOUR
T1 - Expression and purification of bioactive high-purity human midkine in Escherichia coli
A1 - Zhong-hui ZHANG
A1 - Li-juan DU
A1 - Di XIANG
A1 - Shun-ying ZHU
A1 - Ming-yuan WU
A1 - Hui-li LU
A1 - Yan YU
A1 - Wei HAN
J0 - Journal of Zhejiang University Science B
VL - 10
IS - 2
SP - 79
EP - 86
%@ 1673-1581
Y1 - 2009
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B0820385


Abstract: 
Midkine is a heparin-binding growth factor, which plays important roles in the regulation of cell growth and differentiation. The non-tagged recombinant human midkine (rhMK) is therefore required to facilitate its functional studies of this important growth factor. In the present work, rhMK was expressed in Escherichia coli (E. coli) BL21 (DE3). The expression of midkine was efficiently induced by isopropyl-β-D-thiogalactopyranoside (IPTG). After sonication, midkine was recovered in an insoluble form, and was dissolved in guanidine hydrochloride buffer. Renaturation of the denatured protein was carried out in the defined protein refolding buffer, and the refolded protein was purified using S-Sepharose ion-exchange chromatography. The final preparation of the rhMK was greater than 98% pure as measured by sodium dodecylsulfate-polyacrylamid gel electrophoresis (SDS-PAGE) and reverse phase high performance liquid chromatography (RP-HPLC). The purified rhMK enhanced the proliferation of NIH3T3 cells.

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