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《Chinese Journal of Virology》 2006-06
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The Expression of Nucleocapsid Protein of Nipah and Hendra Viruses in Baculovirus and Characterization of Their Reactivity

XIAO Chang~(1,2),XU Xing-ran~1,ZHA Yun-feng~1,ZHANG Yu-jing~2,XUAN Hua~3,TU Chang-chun~(1,2**)(1.Institute of Veterinary Medicine,Academy of Military Medical Science,Changchun 130062,Jilin,China;2.Institute of Animal Husbandry and Veterinary Medicine,Jilin University,Changchun 130062,Jilin,China;3.Institute of Veterinary Medicine,Academy of Guangdong Agricultural Sciences,Guangzhaou 510640, Guangdong,China)  
Nipah virus(NiV) and Hendra virus(HeV),as two novel members of Henipah genus in paramyxoviridae,are the causative agents of fetal Nipah and Hendra zoonoses,which first emerged in 1998 in Malaysia and in 1994 in Australia.These diseases feature the severe pathogenic damages on respiratory system and central nervous system,usually associated with high morbidity and mortality in host animals and the human.In order to prevent their potential outbreak in China,in this study the NiV N and HeV N genes of the viruses were amplified by PCR from the plasmids imported from Australia,and cloned into the pFastBac-Mels-6His vector separately,resulting in the expression in baculovirus system of N protein fused with His-tag at its C terminus.The expression of N proteins in Sf9 cells showed that the highest level of the protein expression could be achieved at 72~96h postinfection,and about 2.3-2.4 mg/L of target proteins could be obtained respectively in large scale expression test.The indirect-ELISA and Western blot results showed that the N proteins expressed could react with rabbit anti-NiV and rabbit anti-HeV sera.The indirect immunofluorescence assay(IFA)using rabbit anti-NiV and rabbit anti-HeV sera could identify the cells expressing N proteins on Sf9 cell monolayers.Taken together,the results showed that the efficient expression of N protein of both NiV and HeV has been achieved and the resulted proteins can be used in future as diagnostic or preventive reagent.
【Fund】: 科技部重大动物疫病防治平台项目(2004BA519A48);; 广东省科技计划项目(2004A20403001)
【CateGory Index】: R373
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