Characterizaiton of virus-like particles prepared by Bac-to-Bac system co-expressing HA and M1 ;protein of highly pathogenic avian influenza H5N1
10.3760/cma.j.issn.0254-5101.2015.04.011
- VernacularTitle:高致病性人禽流感 H5 N1病毒 HA 与 M1蛋白共表达制备新型病毒样颗粒
- Author:
Heng CHEN
;
Jiaming LAN
;
Yang YANG
;
Yuan LIU
;
Jingdong SONG
;
Jianguo QU
;
Jimin GAO
;
Wenjie TAN
- Publication Type:Journal Article
- Keywords:
Influenza virus;
H5N1;
Virus like particles(VLPs);
Baculovirus expression system
- From:Chinese Journal of Microbiology and Immunology
2015;(4):286-291
- CountryChina
- Language:Chinese
-
Abstract:
Objective To express and characterize the virus-like particles( VLPs) of H5 subtype containing of hemagglutinin ( HA ) and matrix 1 ( M1 ) protein by using Baculovirus-insect cells .Methods Full length genes encoding HA protein from the A/Indonesia/05/2005(H5N1) strain and the M1 protein from the A/Anhui/01/2005 ( H5N1 ) strain were cloned into a baculovirus expression vector to construct pFBD-M1-HA.The expression of HA and M1 proteins were detected by Western blot and indirect immunoflu-orescence after the transfection of Spodoptra frugiperda (Sf9) insect cells with recombinant baculovirus.Pu-rified VLPs were analyzed by SDS-PAGE and visualized with transmission electron microscope.The biologi-cal activity of purified VLPs was detected by hemagglutination test.Results The HA and M1 proteins of H5 subtype expressed by baculovirus-insect cells could be self-assembled into the functional mature VLPs.The hemagglutination titer of VLPs was as high as 1024 HAU/50μl.Conclusion The H5 subtype VLPs as pre-pared in this study would pave a way for the development of a candidate recombinant A ( H5) vaccine.
