Linearization of baculovirus DNA enhances the recovery of recombinant virus expression vectors

Nucleic Acids Res. 1990 Oct 11;18(19):5667-72. doi: 10.1093/nar/18.19.5667.


Engineered derivatives of Autographa californica multiple nucleocapsid nuclear polyhedrosis virus (AcMNPV) possessing a unique restriction site provide a source of viral DNA that can be linearized by digestion with a specific endonuclease. Circular or linearized DNA from two such viruses were compared in terms of their infectivity and recombinogenic activities. The linear forms were 15- to 150-fold less infectious than the corresponding circular forms, when transfected into Spodoptera frugiperda cells using the calcium phosphate method. Linear viral DNA was, however, proficient at recombination on co-transfection with an appropriate transfer vector. Up to 30% of the progeny viruses were recombinant, a 10-fold higher fraction of recombinants than was obtained from co-transfections with circular AcMNPV DNA. The isolation of a recombinant baculovirus expression vector from any of the AcMNPV transfer vectors currently in use can thus be facilitated by linearization of the viral DNA at the appropriate location.

MeSH terms

  • Animals
  • Bacterial Proteins*
  • Baculoviridae / genetics*
  • Cells, Cultured
  • DNA, Recombinant / genetics*
  • DNA, Viral / chemistry
  • DNA, Viral / genetics*
  • DNA, Viral / metabolism
  • Deoxyribonucleases, Type II Site-Specific / metabolism
  • Gene Expression*
  • Genes, Viral
  • Genetic Vectors*
  • Moths
  • Plasmids
  • Transfection


  • Bacterial Proteins
  • DNA, Recombinant
  • DNA, Viral
  • BglII endonuclease
  • Deoxyribonucleases, Type II Site-Specific
  • endodeoxyribonuclease SauI