[Expression and identification of sigmaC gene of avian reovirus by baculovirus expression system]

Bing Du Xue Bao. 2011 Jul;27(4):353-7.
[Article in 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.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Baculoviridae / genetics*
  • Capsid Proteins / genetics*
  • Capsid Proteins / metabolism
  • Cell Line
  • Cloning, Molecular
  • Gene Expression*
  • Genetic Vectors / genetics*
  • Genetic Vectors / metabolism
  • Orthoreovirus, Avian / genetics*
  • Orthoreovirus, Avian / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Spodoptera
  • Transfection

Substances

  • Capsid Proteins
  • Recombinant Proteins
  • sigma 1 protein, reovirus