Gene Expression in Mammalian Cells Using BacMam, a Modified Baculovirus System

Methods Mol Biol. 2016:1350:95-116. doi: 10.1007/978-1-4939-3043-2_5.

Abstract

BacMams are modified baculoviruses that contain mammalian expression cassettes for gene delivery and expression in mammalian cells. BacMams have become an integral part of the recombinant mammalian gene expression toolbox in research labs worldwide. Construction of transfer vectors is straightforward using basic molecular biology protocols. Virus generation is based on common methods used with the baculovirus insect cell expression system. BacMam transduction of mammalian cells requires minimal modifications to familiar cell culture methods. This chapter highlights the BacMam transfer vector pHTBV.

Keywords: BacMam; Co-expression; Mammalian expression; Modified baculovirus; Pseudotyping; Transient expression; Viral transduction; Woodchuck hepatitis virus post-transcriptional regulatory element (WPRE).

MeSH terms

  • Animals
  • Baculoviridae / genetics*
  • Baculoviridae / isolation & purification
  • CHO Cells
  • Cloning, Molecular
  • Cricetinae
  • Cricetulus
  • DNA Restriction Enzymes / genetics
  • DNA, Complementary / genetics
  • Gene Expression
  • Genetic Engineering / methods*
  • Genetic Vectors / genetics
  • HEK293 Cells
  • Humans
  • Nucleic Acid Amplification Techniques
  • Sf9 Cells
  • Spodoptera
  • Transformation, Genetic

Substances

  • DNA, Complementary
  • DNA Restriction Enzymes