Mutants of sindbis virus. I. Isolation and partial characterization of 89 new temperature-sensitive mutants

Strauss, E.G.; Lenches, E.M.; Strauss, J.H.

Virology 74(1): 154-168


ISSN/ISBN: 0042-6822
PMID: 62444
DOI: 10.1016/0042-6822(76)90137-9
Accession: 068518999

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More than 100 new temperature-sensitive mutants of Sindbis virus were isolated following mutagenesis with nitrous acid, N-methyl-N'-nitro-N-nitrosoguanidine, 5-azacytidine and 5-fluorouridine. Of these mutants, 36 synthesize at least 60% as much RNA at the nonpermissive temperature as does the parental strain and are designated RNA+; 23 mutants synthesize between 10-60% as much RNA as the parental strain at C and are designated RNA.+-.; 30 mutants make less than 10% as much RNA at C and are called RNA-. The remaining mutants were not tested for RNA incorporation. The thermal stability at C of most of the mutant particles was examined. The majority of the RNA+ mutants is more sensitive to heating at C than the parental HR strain and RNA+ mutations appear to reside primarily in genes coding for the structural proteins. Approximately 20% of either RNA.+-. or RNA- mutants are thermosensitive and these mutations appear to reside primarily in genes coding for the nonstructural proteins. Complementation assays were performed with a number of these mutants and with those of Burge and Pfefferkorn. The existence of 3 complementation groups among the RNA+ mutants which appear to encode the 3 major structural proteins was confirmed; no new complementation groups among RNA+ mutants were identified. A total of 4 complementation groups was identified among the RNA- mutants, Sindbis virus contains at least 7 complementation groups.