Monsanto Company

G.d. Searle & Company

Verne A Luckow

Peter O Olins

Stephen C Lee

Paul Jerome Hippenmeyer

Mark S Leusch

Anne M Rankin

A vaccine is described which comprises an assembly-deficient herpesvirus. The mutant herpesvirus is capable of infecting and undergoing DNA replication in the cells of a susceptible mammal, but is defective in capsid assembly and formation of mature virion particles. The assembly-deficient herpesvirus is avirulent and capable of generating a protective immune response in a vaccinated mammal.

Assembly-deficient herpesvirus vaccine

A method for producing infectious recombinant baculoviruses in bacteria is described. A novel baculovirus shuttle vector (bacmid) was constructed that contains a low-copy-number bacterial replicon, a selectable drug resistance marker, and a preferred attachment site for a site-specific bacterial transposon, inserted into a nonessential locus of the baculovirus genome. This shuttle vector can replicate in E. coli as a plasmid and is stably inherited and structurally stable after many generations of growth. Bacmid DNA isolated from E. coli is infectious when introduced into susceptible lepidopteran insect cells. DNA segments containing a viral promoter driving expression of a foreign gene in insect cells that are flanked by the left and right ends of the site-specific transposon can transpose to the attachment site in the bacmid propagated in E. coli when transposition functions are provided in trans by a helper plasmid. The foreign gene is expressed when the resulting composite bacmid is introduced into insect cells.

Method of producing recombinant eukaryotic viruses in bacteria

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Chesterfield, MO, United States of America

Verne A Luckow