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

• Published in: Nucleic acids research Vol. 18; no. 19; pp. 5667 - 5672
• Main Authors: KITTS, P. A, AYRES, M. D, POSSEE, R. D
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 11.10.1990
• Subjects: Animals; Bacterial Proteins; Baculoviridae - genetics; Biological and medical sciences; Biotechnology; Cells, Cultured; Deoxyribonucleases, Type II Site-Specific - metabolism; DNA, Recombinant - genetics; DNA, Viral - chemistry; DNA, Viral - genetics; DNA, Viral - metabolism; Fundamental and applied biological sciences. Psychology; Gene Expression; Genes, Viral; Genetic engineering; Genetic technics; Genetic Vectors; Methods. Procedures. Technologies; Moths; Plasmids; Transfection; Vectors (cloning, transfer, expression). Insertion sequences and transposons; Recombination; Insecta; Spodoptera frugiperda; Circular DNA; Baculovirus; Gene expression; Nuclear polyhedrosis virus; Virus; Infection; Transfection; Baculoviridae; Arthropoda; Autographa californica; Lepidoptera; Linear shape; Noctuidae; Invertebrata; Vector
• ISSN: 0305-1048; 1362-4962
• 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.