Vaccinia virus-free recovery of vesicular stomatitis virus.
Publication/Presentation Date
10-1-2001
Abstract
The advent of reverse-genetics represents a powerful new approach to elucidate aspects of negative-sense RNA virus replication. The reverse-genetics system established previously for vesicular stomatitis virus (VSV) required four plasmids encoding the nucleoprotein (N), phosphoprotein (P), polymerase (L), and the full-length, anti-genomic RNA. Transcription to yield the antigenomic RNA as well as the N, P, and L, mRNAs was initiated by bacteriophage T7 polymerase expressed from a recombinant Vaccinia virus. In this report, we describe the successful recovery of infectious VSV in the absence of Vaccinia virus. The N, P, and L genes of VSV were inserted downstream of both the T7 promoter and an internal ribosomal entry site (IRES element). T7 polymerase was expressed constitutively from BSR-T7/5 cells. RTPCR was used to confirm that the recovered VSV was derived from transfected DNA. Virion protein profile, CPE in tissue culture, and virus titer of the recombinant VSV were indistinguishable from those of parental VSV. Thus, the need for Vaccinia virus is eliminated with this system, making it an attractive, alternative approach for the recovery of infectious VSV from DNA.
Volume
3
Issue
4
First Page
513
Last Page
517
ISSN
1464-1801
Published In/Presented At
Harty, R. N., Brown, M. E., Hayes, F. P., Wright, N. T., & Schnell, M. J. (2001). Vaccinia virus-free recovery of vesicular stomatitis virus. Journal of molecular microbiology and biotechnology, 3(4), 513–517.
Disciplines
Business Administration, Management, and Operations | Health and Medical Administration | Management Sciences and Quantitative Methods
PubMedID
11545270
Department(s)
Administration and Leadership
Document Type
Article