Expression and Purification of 5'-terminal H Protein of Canine Distemper Virus in E.coli and Analysis its Antigenicity

The aim of this research is was to get high purified 5' terminal H protein of canine distemper virus.One pair of the special primers was designed by Oligo6.0 software according to the hemagglutinin(H) protein gene sequences of canine distemper virus(CDV) published in GenBank.A 388 bp CDV H protein gene fragment was amplified by RT-PCR from the total RNA template CDVs and CDVSD strains.The 5' terminal H protein gene from CDV was cloned into expression vector pET28a(+) and the recombinant plasmid was transformed into E.coli L21(DE3) cells for sequencing,identifying and expressed by IPTG Induced.The constructed pET28a-CDVSD-H5 was successfully expressed at 22kD and purified by Ni-NTA;but the constructed pET28a-CDV3-H5 was not expressed.Western-blotting analysis was shown that could be recognized by CDV positive serum.This research laid the foundation of sero-diagnosis method and genetically engineering vaccine.