Development of baculovirus triple and quadruple expression vectors: co-expression of three or four bluetongue virus proteins and the synthesis of bluetongue virus-like particles in insect cells

• Published in: Nucleic acids research Vol. 21; no. 5; pp. 1219 - 1223
• Main Authors: Belyaev, A.S, Roy, P
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 11-03-1993
• Subjects: Animals; Autographa californica; Baculoviridae - genetics; Baculovirus; Base Sequence; Biological and medical sciences; Biotechnology; Bluetongue virus; Bluetongue virus - genetics; Cell Line; Cloning, Molecular - methods; DNA, Recombinant; DNA, Viral; Electrophoresis, Polyacrylamide Gel; embl/x70469; embl/x70470; Fundamental and applied biological sciences. Psychology; gene transfer; Genetic engineering; Genetic technics; Genetic Vectors; Methods. Procedures. Technologies; Microscopy, Electron; Molecular Sequence Data; Moths; Nuclear polyhedrosis virus; Nucleopolyhedrovirus; Plasmids; Promoter Regions, Genetic; Spodoptera frugiperda; Vectors (cloning, transfer, expression). Insertion sequences and transposons; Viral Structural Proteins - genetics; Viral Structural Proteins - metabolism; Virion - metabolism; Insecta; Spodoptera frugiperda; Recombinant DNA; Baculovirus; Gene expression; Structure gene; Nuclear polyhedrosis virus; Reoviridae; Virus; Transfection; Baculoviridae; Arthropoda; Autographa californica; Lepidoptera; Orbivirus; Noctuidae; Invertebrata; Bluetongue virus
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/21.5.1219

Baculovirus multiple gene transfer vectors pAcAB3 and pAcAB4 have been developed to facilitate the insertion of three or four foreign genes respectively into the Autographa californica nuclear polyhedrosis virus (AcNPV) genome by a single co-transfection experiment. The pAcAB3 vector contains a polyhedrin promoter and two p10 promoters on either side of the polyhedrin promoter but in opposite orientations. The pAcAB4 vector has an additional polyhedrin promoter in opposite orientation to the first copy that is in juxtaposition to the first p10 promoter. Each of these derived vectors (pAcAB3, pAcAB4) have been used for the simultaneous expression of three or four bluetongue virus (BTV) genes respectively. When Spodoptera frugiperda cells were infected with the recombinant virus (AcBT-3/2/7/5) expressing the four major structural genes of BTV, double-capsid, virus-like particles consisting of VP2, VP3, VP5 and VP7 of BTV were assembled.