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

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.