关键词: 汉坦病毒, G2包膜糖蛋白, 昆虫细胞, 间接免疫荧光法

Abstract: Objective To construct hantavirus Z10 envelope glycoprotein gene G2 recombinant and study its application in IFA. Methods Using a pair of primer based on hantavirus Z10 G2 gene sequence. PCR products were obtained by PCR from the plasmid pUCm-Z10M,then digested and cloned into vector pUCm-T which were transformed into E.coli DH5a to construct plasmid pUCm-Z10-G2. The digested product were obtained from plasmid pUCm-Z10-G2 and cloned into Baculovirus transport vestor pFastBacHTa to construct recombinant plasmid pFastBacHTa-Z10-G2 which then were transformed into E.coli DH10Bac. After white E.coli DH10Bac fungus had been cultured and selected for two times,the plasmid pFastBacHTa-Z10-G2 were abstracted again and transfected into insect cells sf9 to express G2 protein,subsequently G2 protein were verified and used to diagnose hantavirus infection by IFA. Results The recombinant baculovirus expressing G2 antigen was constructed and expressed in insect cells. The specific fluorescence in insect cells was observed by IFA. Conclusion The recombinant baculovirus expressing G2 antigen was constructed successfully and expressed in insect cells. The G2 protein as an specific and safe antigen can be used to diagnose hantavirus infection by IFA.