Physical characterization and molecular cloning of the Shope fibroma virus DNA genome

A. Wills, A. M. Delange, C. Gregson, C. Macaulay, Douglas McFadden

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

DNA from several independent strains of Shope fibroma virus, a tumorogenic leporipoxvirus of rabbits, was isolated and analyzed by restriction endonuclease digestion and Southern blotting. The restriction profiles indicated a high degree of sequence conservation among the isolates but blotting under standard stringencies revealed no detectable cross homology with a member of the orthopoxvirus group, vaccinia. The genome of the fibroma virus was calculated to be in excess of 160 kilobases and shown to possess two features analogous to the orthopoxvirus group: (1) the terminal restriction fragments possess covalently closed hairpin structures; and (2) the terminal sequences are present as inverted repeats of greater than 10 kilobases. The terminal 3.6 kilobase BamH1 restriction fragment was cloned in pBR322 after removal of the hairpin structure with mung bean single strand-specific endonuclease and addition of BamHI linkers. SFV sequences within this terminal region were shown, using 32P SFV cloned terminal probe, to have none of the sequence heterogeneity characteristic of vaccinia DNA termini. The remaining 20 internal SFV BamHI restriction fragments were propagated in bacterial plasmids either as intact fragments, or after secondary digestion with HindIII, and together constitute the complete cloned SFV sequence library.

Original languageEnglish (US)
Pages (from-to)403-414
Number of pages12
JournalVirology
Volume130
Issue number2
DOIs
StatePublished - Oct 30 1983
Externally publishedYes

Fingerprint

Rabbit Fibroma Virus
Orthopoxvirus
Vaccinia
Molecular Cloning
Leporipoxvirus
Digestion
Deoxyribonuclease BamHI
Genome
Fibroma
DNA
DNA Restriction Enzymes
Southern Blotting
Libraries
Plasmids
Rabbits
Viruses

ASJC Scopus subject areas

  • Virology

Cite this

Physical characterization and molecular cloning of the Shope fibroma virus DNA genome. / Wills, A.; Delange, A. M.; Gregson, C.; Macaulay, C.; McFadden, Douglas.

In: Virology, Vol. 130, No. 2, 30.10.1983, p. 403-414.

Research output: Contribution to journalArticle

Wills, A. ; Delange, A. M. ; Gregson, C. ; Macaulay, C. ; McFadden, Douglas. / Physical characterization and molecular cloning of the Shope fibroma virus DNA genome. In: Virology. 1983 ; Vol. 130, No. 2. pp. 403-414.
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