A new endonuclease restriction site which is at the locus of a temperature-sensitive mutation in vaccinia virus is associated with true and pseudoreversion

Douglas McFadden, Karim Essani, Samuel Dales

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

A temperature-sensitive mutant of vaccinia IHD-W, designated is 9251, possesses a novel EcoRI restriction endonuclease site in the fragment D of the parental genome. Spontaneous ts+ revertants of ts 9251 fall into two distinct categories: the majority of revertants reacquired the parental EcoRI restriction profile, while one isolate maintained the mutant cleavage site. Using two-dimensional gel analysis of viral polypeptides induced by vaccinia virus in infected cytoplasms, it was observed that the fingerprint of mutant ts 9251 differed from the parental IHD-W in that a single viral protein of a molecular weight of 37,000 daltons migrated to a new isoelectric point. The revertants which had regained the wild-type restriction profile now encoded for a 37K polypeptide identical to that of the wild-type virus while the single revertant which maintained the novel EcoRI site possessed a 37K protein of a charge intermediate between ts 9251 and wild type. We conclude that ts 9251 can revert to the ts+ phenotype either by true reversion at the original mutant locus or by a second independent mutation within the same gene.

Original languageEnglish (US)
Pages (from-to)277-280
Number of pages4
JournalVirology
Volume101
Issue number1
DOIs
StatePublished - Jan 1 1980
Externally publishedYes

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Vaccinia virus
DNA Restriction Enzymes
Deoxyribonuclease EcoRI
Vaccinia
Peptides
Mutation
Temperature
Isoelectric Point
Dermatoglyphics
Viral Proteins
Cytoplasm
Molecular Weight
Gels
Genome
Viruses
Phenotype
Genes
Proteins

ASJC Scopus subject areas

  • Virology

Cite this

A new endonuclease restriction site which is at the locus of a temperature-sensitive mutation in vaccinia virus is associated with true and pseudoreversion. / McFadden, Douglas; Essani, Karim; Dales, Samuel.

In: Virology, Vol. 101, No. 1, 01.01.1980, p. 277-280.

Research output: Contribution to journalArticle

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N2 - A temperature-sensitive mutant of vaccinia IHD-W, designated is 9251, possesses a novel EcoRI restriction endonuclease site in the fragment D of the parental genome. Spontaneous ts+ revertants of ts 9251 fall into two distinct categories: the majority of revertants reacquired the parental EcoRI restriction profile, while one isolate maintained the mutant cleavage site. Using two-dimensional gel analysis of viral polypeptides induced by vaccinia virus in infected cytoplasms, it was observed that the fingerprint of mutant ts 9251 differed from the parental IHD-W in that a single viral protein of a molecular weight of 37,000 daltons migrated to a new isoelectric point. The revertants which had regained the wild-type restriction profile now encoded for a 37K polypeptide identical to that of the wild-type virus while the single revertant which maintained the novel EcoRI site possessed a 37K protein of a charge intermediate between ts 9251 and wild type. We conclude that ts 9251 can revert to the ts+ phenotype either by true reversion at the original mutant locus or by a second independent mutation within the same gene.

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