ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2005, Vol. 36 ›› Issue (4): 376-380.doi:

• 兽医 • Previous Articles     Next Articles

Cloning of σC Gene of Muscovy Duck Reovirus MW9710 and Its Expression in E.coli

ZHANG Yun; OUYANG Sui-dong; LIU Ming;HU Qi-lin;HAN Zhou; LIN Feng-qiang   

  1. 1. National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute,Harbin 150001,China; 2. Fujian Husbandry and Veterinary Research Institute, Fuzhou 350003,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2005-04-25 Published:2005-04-25

Abstract: Minor core protein σC gene of MDRV MW9710 strain was amplified by RT-PCR with the primers designed by the reported muscovy duck reoviruses. The PCR products were cloned into pMD18-T vector and identified by enzyme cutting analysis. The sequence results showed that σC gene is 810 bp. It contains one open reading frame between 280-1089 that encodes a protein of 269 amino acids with a molecular mass of 29.4 ku. Nucleotide analysis by the DNASTAR(6.0) software showed that MDRV MW9710 σC is highly related to the MDRV 89026, with 93.0% homology, but only shared 21%~25% homology to the avian reoviruses. It indicates MDRV is possible to be a new revirus that different from ARV.The DNA fragment of σC was subcloned into prokaryotic expression vector pET32a and the specific fusion protein with molecular weight 50 ku was expressed. Western blotting assay indicated that the polyclonal antibody against DRV could recognize the recombinat σC protein. It also provided technical support for developing an advanced gene engineering vaccine against muscovy duck reovirus.