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| Construction and Immune Efficacy of Recombinant Adenovirus Expressing H Gene of Canine Distemper Virus |
| YUAN Ying-shuo1,2, SONG Fei-fei1,2, LIU Guo-qiang1,2, LIU Ye2, ZHANG Shou-feng2, ZHANG Le-cui1, HU Rong-liang2 |
1. Qingdao Agriculture University, College of Animal Science and Technology, Qingdao 266109, China;
2. Jilin Provincial Key Laboratory of Zoonosis Prevention and Control, and Military Veterinary Research Institute, Academy of Military Medical Sciences, Changchun 130122, China |
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Abstract The recombinant human adenovirus type 5 that expressed hemagglutinin(H) protein gene of canine distemper virus(CDV) be constructed. The shuttle and backbone plasmid were respectively constructed by reverse transcription polymerase chain reaction (RT-PCR) and the restriction enzyme digestion and ligation, then co-transfected into the 293AD cell lines after Pac I linearization respectively. The biological characterization was determined by the electron microscope and restriction enzyme digestion and western blotting analysis and the one-step growth curve, the results proved that the recombinant human adenovirus type 5 that expressed H gene of CDV was constructed correctly. The anti-CDV antibody titers of the dog serum were detected by using neutralization test after a single intramuscular immunization with CDV vaccine strains or recombinants on days 14, 28, 42, 56, 70, 84. The results showed that average anti-CDV antibody titers (up to 2-8) were higher than that of the control group. It implied that the recombinant was constructed correctly and has a good immunogenicity, and it could stimulate an effective level of neutralizing antibodies which reached the lowest level of immune protection against CDV. This work provides a theoretical basis for the research and development of a more efficient distemper live vector vaccine.
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Received: 25 March 2013
Published: 25 January 2014
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