Cloning and Prokaryotic Expession of MD-2 from Buffalo

This study was designed to clone and express buffalo myeloid differentiation-2,and then carry out Western blot for it.RT-PCR was used to clone CDS area of buffalo MD-2,and the PCR products of bMD-2 were cloned into pET-28a,and transformed into E.coli BL21.The bioinformatics analyses were performed.The protein expression was induced by IPTG and analyzed by SDS-PAGE and Western blot.The results showed that the bMD-2 CDS contain a 483 bp ORF,which encodes 161 amino acids,and His-bMD-2 fusion protein was expressed in E.coli BL21(DE3) with molecular weight of 25 ku induced at 37 ℃,6 h by 1 mmol/L IPTG,which indicated that prokaryotic expression vector of buffalo MD-2 was constructed and expressed successfully.