Expression of Cloned Genes Using the Baculovirus Expression System

Cold Spring Harb Protoc. 2020 Jun 1;2020(6):102152. doi: 10.1101/pdb.prot102152.

Abstract

The general strategy of the baculovirus expression system is to infect insect cells with a virus that expresses a foreign protein at a very late stage of infection. Almost all baculovirus expression systems use the procedures for insect cell transfection, baculovirus production, and protein expression given in the main portion of this protocol. This protocol also includes a method that uses molecular biology techniques to produce recombinant baculovirus DNA in E. coli before transfection of insect cells. It is important to quantify the viral titer to achieve optimal and reproducible expression of target proteins. Accordingly, the viral plaque assay is also described here.

MeSH terms

  • Animals
  • Baculoviridae / genetics*
  • Cloning, Molecular / methods*
  • Gene Expression*
  • Genetic Vectors / genetics*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Recombination, Genetic
  • Sf9 Cells
  • Spodoptera
  • Transfection / methods*
  • Viral Plaque Assay / methods

Substances

  • Recombinant Proteins
  • Green Fluorescent Proteins