The expression and bioactivity of S gene in Vero-E6 cells using baculovirus vector
10.3760/cma.j.issn.1003-9279.2011.04.013
- VernacularTitle:杆状病毒为载体的重组汉坦病毒S基因在Vero-E6细胞中的表达及其生物活性
- Author:
Han-Ping ZHU
1
;
Ping-Ping YAO
;
Jie WANG
;
Zhang-Nv YANG
;
Fang XU
;
Rong-Hui XIE
;
Yun ZHANG
;
Zhi-Yong ZHU
Author Information
1. 浙江省疾病预防控制中心
- Keywords:
Hantavirus;
Viral proteins;
Gene expression regulation;
Fluorescent antibody technique,indirect
- From:
Chinese Journal of Experimental and Clinical Virology
2011;25(4):280-282
- CountryChina
- Language:Chinese
-
Abstract:
Objective The S gene of a Hanta Virus (HV) Z10 strain was cloned into a baculovirus shuttle bacmid pDual-CMV contained a CMV promoter to generated a recombinant baculovirus BAC-pDual-CMV-HVS, then the recombinant baculovirus was transfected into Vero-E6 cell. The cells with recombinant baculovirus were applied to the detection of HV antiserum. MethodsTo generate the recombinant baculovirus BAC-pDual-CMV-HVS, the sequence of CMV promoter was obtained from the plasmid pEGFPN1 by PCR, and subsequently cloned to the baculovirus shuttle bacmid pFastBacDUAL resulting the recombinant plasmid pDual-CMV. Then the sequence of HV-S gene was inserted to the plasmid pDualCMV, to generate the plasmid pDual-CMV-HVS. Plasmid pDual-CMV-HVS was transformed into the DH10BAC competent cells to get the recombinant baculovirus BAC-pDual-CMV-HVS. The antigen substrate slides were made by transfecting the recombinant virus into Vero-E6 cells. ResultsThe plasmid pDualCMV-HVS was verified by sequencing. The recombinant virus BAC-pDual-CMV-HVS was generated according to the protocol of the baculovirus and transfected into Vero-E6 cells. The expression of the HV-S gene was verified by positive HV antiserum. Conlusion The recombinant virus were successfully generated and applied to prepare the antigen substrate slides. The antigen substrate slides was conveniently prepared without special equipments, and can be used to detect the antiserum of HV virus.
