Cloning the HSP20 Gene of B.bigemina Isolated in Xinjiang and Construction of the Eukaryotic Expression Plasmid

The special primers were designed by the heat shock protein gene sequence of B.bigemina pubulished in GenBank.and a 699 bp HSP20 gene fragment was amplified by PCR from the whole blood genome in the infected cattle.The PCR product was cloned into a pMD18-T vector,then was sequenced and identified.The 699bp HSP20 gene was then cloned into Eukaryotic vector pcDNA3.1(+)to construct a eukaryotic expression plasmid pcDNA3.1-HSP20 before sequencing and identifying again.Following the pcDNA3.1-HSP20 was transiently expressed in mice injected by invena caudalis,the total RNA was abstructed from murine liver and a 534 bp of bright stripe was found by RT-PCR amplification,suggesting the eukaryotic expression plasmid was successfully constructed.