Replication of Japanese Encephalitis Virus.
Abstract
Gene expression in the flavivirus Japanese encephalitis virus (JEV) was studied by three different approaches. Virus-specific RNA in infected cells was radiolabeled in the presence of actinomycin D, and analyzed by sucrose gradient sedimentation and agarose gel electrophoresis. In addition to the 40S genome RNA and a probable replicative intermediate, two small single-stranded species, designated 15S RNA and 12S RNA, were observed. They were shown by oligonucleotide mapping to be subsets of the 40S RNA and different from each other. Their kinetics of synthesis suggested that they functioned in virus replication, probably as subgenomic mRNAs, and were not merely breakdown products of 40S RNA. Virus-specified proteins were separated from host cell proteins by radioimmune precipitation and analyzed by polyacrylamide gel electrophoresis and fluorography. A library of 15 temperature-sensitive (ts) mutants of JEV were induced by growth of virus in the presence of mutagens. Genetic complementation tests indicated that the mutants represented at least five different gene functions.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 1981
- Accession Number
- ADA127175
Entities
People
- Carol D. Blair
Organizations
- Colorado State University