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Baculovirus Expression System을 이용한 Newcastle Disease Virus 내열성 분리주의 F 단백 유전자 발현
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  • Baculovirus Expression System을 이용한 Newcastle Disease Virus 내열성 분리주의 F 단백 유전자 발현
저자명
장경수,김지영,김석,김태용,송희종,전무형,Chang. Kyung-Soo,Kim. Ji-Young,Kim. Suk,Kim. Tae-Yong,Song. Hee-Jong,Jun. Moo-Hyung
간행물명
Journal of bacteriology and virology : JBV
권/호정보
2001년|31권 2호|pp.163-174 (12 pages)
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대한미생물학회
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

The gene (1,707 bp) encoding fusion (F) protein of CBP-1 strain, a heat-stable Newcastle disease virus (NDV) isolated from the diseased pheasant in Korea was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the clone of pVL-NDF inserted with NDV F gene was constructed. Using Baculo Gold Transfection kit, pVL-NDF plasmid and linear baculovirus DNA were cotransfected into Sf-9 cells, and F gene in NDVF recombinant baculovirus was identified by restriction enzyme cleavage patterns and Southern blot hybridization. The Sf-9 cells infected with the NDVF recombinant baculovirus showed the typical cytopathic effects with syncytial cell formation. The properties of the expressed F protein were examined by indirect fluorescent assay, immunocytochemistry and indirect dot-immunoassay. By SDS-PAGE, the expressed F protein with the size of 55 kDa was detected in the cells infected for 5 days with 0.125 to 4.0 moi of the recombinant baculovirus and in the cells infected with 2 moi of the recombinant baculovirus for 1 to 6 days. The guinea pigs inoculated with the mixtures of the infected Sf-9 cell cultures and adjuvants showed significant immunoreactivity, as measured by indirect enzyme-linked immunosorbent assay.