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Báo cáo khoa học: " Expression of Open Reading Frame 5 Protein of Porcine Reproductive and Respiratory Syndrome Virus Using Semliki Forest Virus Expression System"

Tuyển tập các báo cáo nghiên cứu khoa học quốc tế về bệnh thú y đề tài: Expression of Open Reading Frame 5 Protein of Porcine Reproductive and Respiratory Syndrome Virus Using Semliki Forest Virus Expression System | J. Vet. Sci. 2002 3 1 13-18 JOURNAL OF Veterinary Science Expression of Open Reading Frame 5 Protein of Porcine Reproductive and Respiratory Syndrome Virus Using Semliki Forest Virus Expression System Hae-Sun Jung In-Wook Hwang Su-Mi Kim Chul-Joong Kim Kwang-Soon Shin and Hyun-Soo Kim College of Veterinary Medicine Chungnam National University Daejeon 305-764 Korea ABSTRACT The ORF5 gene encodes a major envelope glycoprotein GP5 which is one of the three major proteins of porcine reproductive and respiratory syndrome virus PRRSV . The GP5 protein has been known to be a 24.5-26kDa N-glycosylated envelope protein. The GP5 is involved in inducing neutralizing antibodies. For this reason the GP5 is primary candidate for the PRRSV subunit vaccine. To produce the native form of GP5 in mammalian cells we have cloned the ORF5 gene from PRRSV CNV-1 into the Semliki Forest virus SFV -based expression vector resulting in recombinant pSFV-ORF5. By the infection with recombinant pSFV-ORF5 to BHK-21 cells the GP5 expression was confirmed by immunocytochemistry and immunoblotting assay. The recombinant virus particle harboring ORF5 gene was infectious to BHK-21 and MARC-145. The RNA synthesis and expression of GP5 in the infected cell was also confirmed by RT-PCR. Key words PRRS virus ORF5 gene GP5 Semliki Forest virus-based expression vector recombinant SFV particle. Introduction Porcine reproductive and respiratory syndrome PRRS is a disease found in swine farms worldwide and it is characterized by reproductive failures such as late-term abortions in sows and by respiratory illness and high mortality in young pig1 2 6 7 8 10 1 PRRSV are related to those of a group of small enveloped positive-strand RNA viruses including murine lactate dehydrogenase elevating virus LDV equine arteritis virus EAV and simian haemorrhagic fever virus which are presently classified Corresponding author Dr. Hyun-Soo Kim Division of Preventive Medicine College of Veterinary Medicine Chungnam National