Cloning and Expression of the H Protein Fragment of Canine distemper virus Yunnan Strain

Two pairs of primers were designed and synthesized based on the Haemgglutinin(H) protein gene sequerce of the Onderstepoort strain of Canine distemper virus(CDV).The templates were produced from the reverse transcription reaction,of which total RNA was isolated from vero cell infected Yunnan isolate of CDV.The H1 and H2 gene fragment were amplified by polymerase chain reaction(PCR).The 939 bp and 920 bp fragment of the PCR products digested with BamHI and PstI were cloned into the expression plasmid vector PET-28a(+),respectively.The recombinant were transformed into the BL21(DE3)plysS and induced to express by 1.0 mmol/L IPTG at 35 ℃.The expressed products of 34 ku were identified by SDS-PAGE and Western blot with CDV Onderstepoort antiserum.The results revealed that the expressee fusion protein in vitro had the critical antigenit epitopes of H gene of CDV.