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Expression, Purification of N-terminus Fusion Human MCM7 and a Study on Its Interaction with AR |
1.National Glycoengineering Research Center, Shandong University, Jinan 250012, China
2.Central Hospital of Zaozhuang Coal Mining Group Co. Ltd, Zaozhuang 277011, China |
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Abstract 744 bp of N- terminal of mcm7 gene was amplified by PCR and was cloned into prokaryotic expressing vector pGEX-5x-3 to construct pGEX-5x-3/MCM7N. A batch of E.coli BL-21 were transformed with pGEX-5x-3 and pGEX-5x-3/MCM7N, respectively. GST protein and GST-MCM7N fusion protein were obtained from the recombinant E. coli BL-21 after IPTG induction and were purified with Glutathione Sepharose 4B. The fusion proteins with 54 kDa molecular weight were specifically recognized by both anti-GST antibody and anti-MCM7 antibody in Western blotting analyses. GST pull-down analysis showed that GST-MCM7N fusion proteins interacted directly with androgen receptor (AR) protein after GST-MCM7N incubated with prostate cancer LNCaP cell lysate and His-AR fusion protein, respectively. The result demonstrated that the N- terminal of MCM7 protein can interact directly with AR in vitro.
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Received: 12 June 2009
Published: 26 February 2010
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Corresponding Authors:
Yi-kang Shi
E-mail: shiyikang@yahoo.com
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