Project Number: 3622-22000-029-00
Project Type: Appropriated

Start Date: May 25, 2005
End Date: May 24, 2010

Objective:
Genetically engineer specific fluorescent protein genes into baculoviruses that will be expressed in the envelopes of budded virus and occlusion bodies to serve as barriers against ultraviolet inactivation. Incorporate specific proteins into polyhedra (occlusion bodies) of baculoviruses by a genetic fusion process that would lead to early mortality or inhibit feeding of pest insects. Examine the basis of baculovirus resistance at the cellular level employing cell culture methodology in order to better understand the nature of baculovirus host range.

Approach:
This proposal employs a molecular approach whereby baculoviruses will be stabilized against inactivation by ultraviolet light (UV) as well as to make them faster acting. To achieve these goals, several fluorescent protein genes will be engineered into baculovirus genomes which when expressed in the envelopes of the viral particles and occlusion bodies will provide protection against inactivation by UV light. This methodology is designed to reduce damage to the viral DNA of baculoviruses. To make the bacuoviruses faster acting, specific proteins will be incorporated into occlusion bodies of baculoviruses by a genetic fusion process that will lead to early mortality or inhibit feeding by pest insects since viral replication in the host is not a pre requisite for elaboration of the active proteins. This research project will also examine the nature of baculovirus resistance at the cellular level employing insect cell lines that are refractile to baculoviruses. Recombinants with fluorescent markers under various promoters will elucidate where the block occurs and thus provide knowledge that could be valuable in understanding viral host range. Recombinant research to be conducted in this laboratory has been approved at BSL-1 (#90-78) on November 24, 2004 by Environmental Health & Safety, University of Missouri-Columbia.