Use of a hybrid infectivity assay to analyze primary transcription of temperature-sensitive mutants of the New Jersey serotype of vesicular stomatitis virus

Abstract

A hybrid infectivity assay specific for primary transcription was developed to analyze the production of functional mRNAs by vesicular stomatitis virus. A template prepared from wild-type virions of the New Jersey serotype of vesicular stomatitis virus was reconstituted with RNA polymerase proteins from the wild type or temperature-sensitive mutants, and the in vivo temperature sensitivity of the polymerase was determined by infectivity assay. The data demonstrate that the New Jersey temperature-sensitive mutants A1 and E1 have non-temperature-sensitive transcriptases, whereas the B1 and F1 mutants both have temperature-sensitive L proteins which are defective in primary transcription.