China Animal Husbandry & Veterinary Medicine ›› 2012, Vol. 39 ›› Issue (2): 20-24.

• 生物技术 • Previous Articles     Next Articles

Cloning and Prokaryotic Expession of MD-2 from Chinese Holstein Dairy Cow

JIAO Han-wei1, WANG Hong-mei2, LIU Xiao2, DU Li1, LEI Ming1, ZHANG Dong-lin1, HAO Yong-chang1, CHENG Ying1, HE Hong-bin2, WANG Feng-yang1   

  1. 1. Hainan Key Laboratory of Tropical Animal Reproduction & Breeding and Epidemic Disease Research (Construction Period),Animal Genetic Engineering Key Laboratory of Haikou,College of Agriculture,Hainan University,Haikou 570228,China;2. Dairy Cattle Research Centre,Shandong Acdemy of Agriculture Science,Jinan 250183,China
  • Received:2011-07-01 Revised:1900-01-01 Online:2012-02-20 Published:2012-02-20

Abstract: In order to clone,express and identify myeloid differentiation-2 of Holstein dairy cow,total RNA was isolated from Chinese Holstein dairy cow,RT-PCR was used to amplify MD-2 cDNA. The PCR products were and cloned into pMD20-T vector and the recombinant plasmid was confirmed by PCR,endonuclease digestion. MD-2 was cloned into pET28a vector to construct the pET28a-MD-2,protein expression was induced by IPTG and analyzed by SDS-PAGE and Western blotting. The results showed that the MD-2 gene contain a 483 bp ORF encoding 161 amino acids; His-MD-2 fusion protein was expressed in E.coli BL21(DE3) with molecular weight of 25 ku after induced by IPTG.

Key words: Chinese Holstein dairy cow; MD-2; cloning; prokaryotic expression

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