Expression and identification of sigmaC gene of avian reovirus by baculovirus expression system.
- Author:
Mei-Yu SUN
1
;
Li-Ting QIN
;
Yu-Long GAO
;
Xiao-Le QI
;
Hong-Lei GAO
;
Yong-Qiang WANG
;
Xiao-Mei WANG
Author Information
1. Division of Avian Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy Agricultural Sciences, Harbin 150001, China. meiyu01@126.com
- Publication Type:Journal Article
- MeSH:
Animals;
Baculoviridae;
genetics;
Capsid Proteins;
genetics;
metabolism;
Cell Line;
Cloning, Molecular;
Gene Expression;
Genetic Vectors;
genetics;
metabolism;
Orthoreovirus, Avian;
genetics;
metabolism;
Recombinant Proteins;
genetics;
metabolism;
Spodoptera;
Transfection
- From:
Chinese Journal of Virology
2011;27(4):353-357
- CountryChina
- Language:Chinese
-
Abstract:
Recombinant baculovirus containing sigmaC gene of Avian reovirus was constructed using Bac-To-Bac Baculovirus expression system, and recombinant sigmaC protein was expressed by infecting the sf9 cell with recombinant baculovirus. Firstly, sigmaC gene of Avian reovirus was cloned and inserted into donor plasmid pFastBacHTA to obtain recombinant donor plasmid pFsigmaC. Plasmid pFsigmaC was transformed into E. coli DH10Bac for integration into bacmid vector and the recombinant bacmid plasmid BacmidsigmaC was obtained. Recombinant baculovirus rBacsigmaC was obtained by transfection of the sf9 cells with BacmidsigmaC. Western blot and indirect immunofluorescence assay (IFA) were carried and the results showed that the recombinant sigmaC protein with 37 kDa molecular weight was expressed successfully.
