[Use of a novel baculovirus vector to express nucleoprotein gene of Crimean-Congo hemorrhagic fever virus in both insect and mammalian cells].
OBJECTIVE
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.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Ma
Benjiang
B
Department of Hemorrhagic Fever and Arboviruses, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Hang
Changshou
C
Zhao
Yun
Y
Wang
Shiwen
S
Xie
Yanxiang
Y
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D015870:Gene Expression; D005814:Genes, Viral / Q000235:genetics*; D005822:Genetic Vectors; D003225:Hemorrhagic Fever Virus, Crimean-Congo / Q000235:genetics*; D014162:Transfection; D014758:Viral Core Proteins / Q000235:genetics*
D014758:Viral Core Proteins