Expression of recombinant VP_2 gene in insect sf9 cells and screening of clinical specimens
10.3760/cma.j.issn.1003-9279.2009.06.009
- VernacularTitle:博卡病毒VP_2基因在昆虫细胞sf9中表达及临床标本筛查
- Author:
Ling-Fang TENG
1
;
Feng LIN
;
Mei-Yun ZHENG
;
Chang-Hua ZHENG
;
Feng WU
;
Ai-Ping ZENG
;
En-Pei HUANG
;
Yi-Han MO
;
Min-Qiao ZHENG
;
Xu-Yang LI
;
Jian-Yi HOU
Author Information
1. 温州医学院附属温岭医院
- Keywords:
Parvovifidae;
Gene expression profding;
lmmunoblotting
- From:
Chinese Journal of Experimental and Clinical Virology
2009;23(6):427-429
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone and express VP_2 gene from HBoV,and the expressed VP_2 protein was as the antigen in order to detect serum from children in Wenling area with lower respiratory tract infections.Methods The VP_2 gene was reeombined with the genome of Baculovirus,which infected the insect cell.The fusion protein with HA tag was applied to confirm the specificity of expressed protein.Furthermore,the recombinant protein was observed using electron microscopy.The 176 serum from children in Wenling area with lower respiratory tract infections was screened using Western blot.Results The expressed VP_2 protein was more than 60% in total proteins from insect cell,and MWt about 60 ×10~3.The virus-like particle(VLP)Was observed using electron microscopy,and size about 20nm.The 176 serum from children in wenling area with lower respiratory tract infections was screened using Western blot.The HBoV positive rate Was 2.28%(4/176).Conclusion The VP_2 protein from human bocavims was expressed in insect cell successfully.Through HA tag the VP_2 protein wag specific,and then the assay using SDS-PAGE with Western blot could detect and screen the antibody in serum from children with lower respiratory tract infections rapidly and accurately.
