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바큘로바이러스를 이용한 돼지 2형 써코바이러스 유사입자 발현
김하현 ( Ha Hyun Kim ),박건용 ( Geon Yong Bak ),박준규 ( Jun Gyu Park ),김현정 ( Hyun Jeong Kim ),권형준 ( Hyoung Jun Kwon ),박수진 ( Su Jin Park ),정영주 ( Young Ju Jeong ),김채현 ( Chae Hyeon Kim ),( Myra Hosmillo ),( Mia 한국수의공중보건학회 2010 예방수의학회지 Vol.34 No.3
Porcine circovirus type 2 (PCV2) is associated with porcine circovirus diseases (PCVD), of which postweaning multisystemic wasting syndrome is considered to cause considerable economic losses in pig industry worldwide. As the virus-like particle (VLP) is a highly effective type of subunit vaccine and has unique advantages in terms of safety and immunogenicity, this study aimed to develop PCV2-like particles, which matched currently circulating Korean PCV2 and were applicable as vaccines. The ORF2 genes encoding PCV2 capsid protein were amplified from the PCV2 subgroup 1A/B Korean C275 isolate and the subgroup 2E C94 isolate by PCR assay with primer pair specific to PCV2 ORF2 gene, and were cloned into baculovirus transfer vector. Recombinant baculovirus was generated by cotransfection with the transfer vector and linear baculovirus DNA into the Sf9 cells, and then by plaque purification. Expression of PCV2 capsid protein was determined by the indirect immunofluorescence and Western blotting assays, and electron microscopy. By both immunological assays, PCV2 capsid antigen was detected in the Sf9 cells infected with the recombinant baculoviruses. The formation of empty virus particles, characteristic of VLP, was detected in the lysate of Sf9 cells infected with the recombinant baculoviruses by negative electron microscopy. From these results, VLPs of two genogroups of PCV2 were successfully expressed and generated in a baculovirus expression system. It is expected that the expressed VLPs of two genotypic groups can be used for control of PCV2 infection as good vaccine candidates.