A hybrid baculovirus-bacteriophage T7 transient expression system

Biotechnology (N Y). 1995 Mar;13(3):261-4. doi: 10.1038/nbt0395-261.

Abstract

A hybrid recombinant baculovirus-bacteriophage T7 expression system was developed for transient expression in insect cells of plasmids with foreign genes provided with a T7 promoter. The coding sequence for T7 RNA polymerase, with or without a nuclear localization signal, was inserted into the genome of Autographa californica nuclear polyhedrosis virus. Recombinant viruses stably expressed T7 RNA polymerase in insect cells. Upon transfection of infected insect cells with plasmids containing the genes for chloramphenicol acetyltransferase (CAT), the hepatitis B virus precore-, core- or e- antigens under control of the T7 promoter, transient expression of these genes was detected by ELISA. The results obtained indicate that this baculovirus/T7 system provides a simple and widely applicable tool for transient gene expression studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacteriophage T7 / genetics*
  • Baculoviridae / genetics*
  • Cell Nucleus / enzymology
  • Chloramphenicol O-Acetyltransferase / genetics
  • Cytoplasm / enzymology
  • DNA, Recombinant
  • DNA-Directed RNA Polymerases / genetics*
  • Gene Expression*
  • Hepatitis B Core Antigens / genetics
  • Hepatitis B e Antigens / genetics
  • Plasmids / genetics
  • Promoter Regions, Genetic
  • Protein Sorting Signals / genetics
  • Recombinant Fusion Proteins
  • Spodoptera / metabolism
  • Spodoptera / ultrastructure
  • Transfection
  • Viral Proteins

Substances

  • DNA, Recombinant
  • Hepatitis B Core Antigens
  • Hepatitis B e Antigens
  • Protein Sorting Signals
  • Recombinant Fusion Proteins
  • Viral Proteins
  • Chloramphenicol O-Acetyltransferase
  • bacteriophage T7 RNA polymerase
  • DNA-Directed RNA Polymerases