ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2006, Vol. 37 ›› Issue (4): 412-416.doi:

• 研究简报 • Previous Articles    

Construction, Expression of Gene Vaccine Vectors of Canine Coronavirus and Its’ Immunogenicity

QIAO Jun;XIA Xian-zhu;YANG Song-tao; HAO Feng; GAO Yu-wei;ZHENG Xiao-yi;HUANG Geng   

  1. 1. Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun 130062,China; 2. 791 Police Dog Base,Harbin 150078,China; 3. Department of Animal Science and Technology, Tarim University, Alar 843300,China;4.College of Life Science,Jilin University,Changchun 130012,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2006-04-25 Published:2006-04-25

Abstract: In this experiment, three recombinant expression vectors based on pVAX1 were successfully constructed. According to the sequences of spike (S) gene, membrane protein (M) gene and nucleoprotein (N) gene of CCV Insavc-1 strain, three pairs of specific primers were designed and used to amplify S, M and N genes of DXMV strain. The PCR products were cloned into pGEM-T. Then the recombinant pTS, pTM and pTN were used for sequencing. After sequenced, three immunogenicial genes were subcloned into pVAX1, respectively, which named pVAXS, pVAXM and pVAXN. The three constructed expression vectors were transfected into MDCK cell. Transcriptions of interesting genes were detected by RT-PCR. The expression of target proteins was detected by indirect ELISA. The results showed that target genes can be transcripted after 36 hours , and target proteins can be detected after 72 hours of transformation. Immunization in dogs indicated that the three expression vectors can efficiently induce the specific cellular and humoral immunoresponse, which laid a solid foundation for the further studies of new vaccines of CCV.