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Virologica Sinica, 19 (5) : 487-489, 2004
Research Article
Cloning and Expression of the H Protein Gene Fragment of Canine Distemper Virus Nanjing Strain
Key Lab of Animal Diease Diagnostic and Immunology, Ministry of Agriculture, Nanjing Agriculture University, Nanjing 210095,China
 Correspondence:
(1091.69KB)  
Abstract
One pair of primers was designed and synthesized based on the Haemagglutinin (H) protein gene Sequercein the Onderstepoort strain of Canine distemper virus(CDV). The templates were produced from the reverse transcription reaction, total RNA was isolated from the Vero cells infected with the Nanjing isolate of CDV (CDV NJ-15). The full-length H gene fragment was amplified by polymerase chain reaction (PCR). Digested with EcoR I and Sal I, a 1058bp fragment of the PCR product was cloned into the expression plasmid vector pET-28b (+).The recombinant was transformed into the RosettaTM and induced to express by 1.0m mol/L IPTG at 37℃.The expression product was identified by SDS-PAGE and found to be 38kDa as expected and confirmed by Western blotting with CDV-Onderstepoort rabbit antiserum. The results revealed that the expressed fusion protein in vitro had the critical antigenitic epitopes of H gene of CDV.
  Published online: 20 Oct 2004
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