Scientia Agricultura Sinica ›› 2007, Vol. 40 ›› Issue (2): 373-378 .doi: 10.3864/j.issn.0578-1752.at-2006-7386

• ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT • Previous Articles     Next Articles

Construction of Prokaryotic Expression Vector of Mouse Nanog Gene and its Expression

  

  1. 西北农林科技大学国家干细胞工程技术研究中心陕西分中心
  • Received:2006-04-07 Revised:2006-05-23 Online:2007-02-10 Published:2007-02-10

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Abstract: A pair of primers were designed according to enzyme digestion sites in pGEX-KG and the Nanog gene sequence published by Genbank. The DNA fragment of 918bp was amplified by PCR from the pNA992 recombinant plasmid with Nanog gene, then cloned into pGEX-KG and transformed into the host E.coli strain TGⅠ. The fragment was conformed to the original sequence. It indicated that fusion expression vector pGEX-KG-Nanog was constructed．The recombinant plasmid was taken and transformed into BL21(DE3) for expression. Induced by IPTG at 37℃, the expression product of Nanog gene was identified by SDS-PAGE. The results revealed that Nanog protein had been expressed successfully in the form of inclusion bodies, The molecular weight is 63kDa.Meanwhile,The optimum condition of Nanog protein expression was induced with 0.8mmol/L IPTG for 5h.

Key words: Nanog gene, Prokaryotic expression, GST fusion protein, Mouse

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