Expression of envelope protein of Zika virus in baculovirus expression system
10.3760/cma.j.issn.1003-9279.2017.06.018
- VernacularTitle: 寨卡病毒E蛋白在重组杆状病毒中的表达
- Author:
Hanchun GAO
1
;
Lihong YAO
;
Chao WANG
;
Lishu ZHENG
Author Information
1. National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China
- Publication Type:Journal Article
- Keywords:
Zika virus;
Envelope protein;
Baculovirus;
sf9 cells
- From:
Chinese Journal of Experimental and Clinical Virology
2017;31(6):562-565
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To express envelope protein of ZIKA virus in baculovirus expression system.
Methods:Full-length E gene of ZIKA virus was obtained by DNA synthesis and inserted into vector pFastBac1. The constructed recombinant baculovirus transfer vector pFB1-E was transformed to competent DH10Bac cells. The obtained skeleton plasmid rBacmid-E was transfected to sf9 cells, and the constructed recombinant baculovirus rBac-E was determined for titer, for insertion of E gene by PCR, and for expression of E protein by IFA and Western blotting.
Results:PCR proved that skeleton plasmid rBacmid-E was constructed correctly. The titer of rBac-E of passage 3 was 2.58×105pfu/ml. The genome of infected cells virus was extracted, the gene band at length of 3 830 bp was observed after PCR amplification. Indirect immunofluorescence of the infected cells showed the specific green fluorescence, 55×103specific band was determined by Western blotting identification in the cell pellet of the infected recombinant baculovirus rBac-E.
Conclusions:The recombinant baculovirus with E gene of ZIKA virus was successfully constructed, which laid a foundation of further study on the function of E protein and the vaccine of ZIKA virus.
