Ma Benjiang, Hang Changshou, Zhao Yun, Wang Shiwen, Xie Yanxiang
Department of Hemorrhagic Fever and Arboviruses, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2002 Sep;16(3):249-52.
To construct a novel baculovirus vector which is capable of promoting the high-yield expression of foreign gene in mammalian cells and to express by this vector the nucleoprotein (NP) gene of Crimean-Congo hemorrhagic fever virus (CCHFV) Chinese isolate (Xinjiang hemorrhagic fever virus, XHFV) BA88166 in insect and Vero cells.
Human cytomegalovirus (CMV) immediate early (IE) promoter was ligated to the baculovirus vector pFastBac1 downstream of the polyhedrin promoter to give rise to the novel vector pCB1. XHFV NP gene was cloned to this vector and was well expressed in COS-7 cells and Vero cells by means of recombinant plasmid transfection and baculovirus infection.
The XHFV NP gene in vector pCB1 could be well expressed in mammalian cells. Vero cells infected with recombinant baculovirus harboring NP gene could be employed as antigens to detect XHF serum specimens whose results were in good correlation with those of ELISA and in parallel with clinical diagnoses.
This novel baculovirus vector is able to express the foreign gene efficiently in both insect and mammalian cells, which provides not only the convenient diagnostic antigens but also the potential for developing recombinant virus vaccines and gene therapies.
构建一种新型杆状病毒载体，使其能够促进外源基因在哺乳动物细胞中的高产表达，并利用该载体在昆虫细胞和Vero细胞中表达克里米亚-刚果出血热病毒（CCHFV）中国分离株（新疆出血热病毒，XHFV）BA88166的核蛋白（NP）基因。
将人巨细胞病毒（CMV）立即早期（IE）启动子连接到杆状病毒载体pFastBac1的多角体蛋白启动子下游，构建新型载体pCB1。将XHFV NP基因克隆到该载体中，通过重组质粒转染和杆状病毒感染在COS-7细胞和Vero细胞中良好表达。
载体pCB1中的XHFV NP基因在哺乳动物细胞中能良好表达。用携带NP基因的重组杆状病毒感染的Vero细胞可作为抗原检测XHF血清标本，结果与ELISA法高度相关且与临床诊断相符。
这种新型杆状病毒载体能够在昆虫细胞和哺乳动物细胞中高效表达外源基因，不仅提供了便捷的诊断抗原，还为开发重组病毒疫苗和基因治疗提供了潜力。
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