Stable Pre-Occluded Virus Particle For Use in Recombinant Protein Production and Pesticides
Patents No. 5,593,669 and 6,090,379
Issued: January 14, 1997 and July 18, 2000
Inventor: H.Alan Wood, Ithaca, NY
A method of infecting insects is disclosed; the method utilizes a form of a baculovirus which is highly efficient at establishing infection and is normally destined to become occluded within the polyhedrin or granulin -- Pre-occluded Virus (POV). Specifically, the POV as derived from a polyhedrin-minus or granulin-minus (lacking a functional polyhedrin or granulin gene) baculovirus is fed to insect larvae per os resulting in high infection rates. The stabilization and use of the POV form of polyhedrin-minus baculoviruses as an insecticide is also disclosed.
The inventor of the subject invention shows in the subject application that the formation and accumulation of POV occurs during the replication of a polyhedrin-minus baculoviruses (Figure 1), and that these POV can be stabilized and are infectious per os using the neonate droplet feeding method (Hughes, P.R., Wood, H.A., 1981, "A synchronous peroral technique for the bioassay insect viruses", J. Invert. Pathol. 37:154-159; and Hughes, P.R., Van Beek, N.A.M., Wood, H.A., 1986, "A modified droplet feeding method for rapid assay of Bacillus thuringiensis and baculoviruses in noctuid larvae", J. Invert. Pathol. 48:187-192). Thus, this invention represents a simple solution to a problem in a manner which is contrary to the teachings and expectations of the prior art.
A method of infecting insects is disclosed; the method utilizes a form of a baculovirus which is highly efficient at establishing infection and is normally destined to become occluded within the polyhedrin or granulin -- Pre-occluded virus particles (POV). Specifically, the POV produced during replication of polyhedrin-minus (genetic alteration resulting in nonfunctional polyhedrin or granulin gene) baculovirus is highly infectious to insect larvae when administered per os. The discovery of a stable POV form of polyhedrin-minus baculoviruses was essential to our invention of the novel method of infecting insect larvae per os using the POV form of polyhedrin-minus baculovirus, and as such is also described and taught in the specifications.
As stated above, stabilizing the POV without losing infectivity is very important. The inventor was genuinely surprised to discover that freeze-drying had little effect on the infectivity of the POV produced in larvae. Therefore, a stable and highly infectious POV preparation is one of the many important discoveries of the present invention.
It is therefore an object of this invention to disclose a novel method of infecting insects per os with POV formed during the replication of polyhedrin-minus or granulin-minus baculoviruses.
It is another object of this invention to disclose a novel method of making POV derived from polyhedrin-minus or granulin-minus baculovirus during the course of replication in insect tissue.
It is yet another object of this invention to disclose a novel pesticide; wherein said POV derived from a polyhedrin-minus or granulin-minus baculovirus is the active agent of said pesticide.
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