Physical mapping, expression, and genetic manipulation of the Autographa Californica nuclear polyhedrosis virus genome

Abstract

A translational map of the Autographa californica nuclear polyhedrosis virus (AcMNPV) genome and the approximate location of 19 genes were determined. The gene for AcMNPV polyhedrin, the major component of the viral occlusions, was encoded for by a contiguous DNA sequence located about 4000 to 5200 bases to the right of the origin of the AcMNPC physical map. The DNA sequence for the polyhedrin gene was determined. Deletions were introduced in the AcMNPV polyhedrin gene and the mutant genes were transferred to viral genomes. The resultant viral mutants were infectious in cultured cells and the host organism, but no viral occlusions formed during infection. Using specially constructed plasmids, the protein-coding sequences for human beta interferon were linked to the AcMNPV polyhedrin promoter. The polyhedrin-interferon hybrid genes were transferred to infectious AcMNPV recombinant viruses which were used to infect cultured insect cells. Biologically active interferon was produced, glycosylated, and secreted from infected cells. These results demonstrate that AcMNPV would be suitable for use as a eucaryotic expression vector for the production of products from cloned genes.