The objective of this project is to develop cloning and expression vectors from nuclear polyhedrosis viruses (NPV) with capabilities for efficiently and simultaneously producing one of several genes through the isolation and engineering of at least two new polyhedrin gene promoters in infected insect cell cultures. Alternative plasmid transfer vectors with increased expression capabilities will be constructed in Phase 1. These transfer vectors will be used in the construction of viral expression vectors capable of producing more than one gene product at one time under the control of polyhedrin promoters.Phase ll will concentrate on the expression of specified genes in the vectors, optimization of large-scale cultures of vector-infected insect cells, and purification procedures for the gene products. Preliminary studies have demonstrated that this expression system allows posttranslational modification, glycosylation, and efficient secretion of gene products from infected cells. During the course of this study investigators will also complete a detailed cost analysis comparing this system with classical bacterial and yeast expression systems. The new vectors developed in this study will enhance the commercial feasibility of NPV-infected cell cultures as efficient expression systems for many gene products that could be marketed as research and clinically therapeutic reagents.Institute Of Allergy And Infectious Diseases
