Flavivirus RNA Replication: Essential Viral Functions as Targets for Antiviral Therapeutics
Abstract
The objective of this contract research is to gain a detailed understanding of the molecular events involved in flavivirus RNA replication, with the ultimate aim being the identification of inhibitors of the process. We established yellow fever virus (YFV) as our study system. Necessary virologic, molecular biologic, and immunologic reagents have been obtained or were generated during this first year of work. Methodologies for detection, identification, and analysis of YFV polypeptides and RNA species are in place. Select YFV protein coding regions were engineered into transcription expression plasmids and baculovirus transfer vectors. Evaluation of gene expression of the former constructs was performed in a mammalian cell transient transfection system. Several of the latter vectors were used to construct recombinant baculoviruses. Protein expression by two of them (bacNS3 and bacNS5) was established. The NS5 protein produced in bacNS5-infected Sf9 cells have been purified. A specific, reproducible, and quantifiable assay for RNA-dependent RNA polymerase (RDRP) activity involving a subcellular fraction derived from YFV- infected cells has been established.
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 30, 1991
- Accession Number
- ADB157113
Entities
People
- Joann A. Suzich
- Marc S. Collett