Cloning, restriction endonuclease mapping and partial sequence analysis of the genome of human herpesvirus 7 strain JI

Vivian R. Ruvolo, Zwi Berneman, Paola Secchiero, John Nicholas

Research output: Contribution to journalArticlepeer-review

Abstract

Human herpesvirus 7 (HHV-7) is a recently isolated herpesvirus that has been shown to be related to human cytomegalovirus and human herpesvirus 6 and to be a member of the betaherpesvirus subgroup. Here we report the cloning, restriction endonuclease mapping and partial sequence analysis of HHV-7 strain JI DNA. Virus particles were obtained from the supernatant of infected SupT1 cells, the DNA isolated by proteinase K treatment-phenol extraction, and full-length viral DNA was purified and isolated on a pulsed-field gel. Aliquots of this highly purified material were treated in the following ways: (i) sonicated and end-repaired to create short randomly sheared fragments for cloning into M13mpl8-Smal vector DNA; (ii) cut with EcoRI for cloning into EcoRI-cut λZAPII or λDASHII vectors; (iii) cut with BamHI for cloning into BamHI-cut λZAP-Express or λDASHII vectors. Partial nucleotide sequencing of the M13 clones followed by detection of open reading frames and their translation allowed the identification of homologues through FASTA searches of the database. Relevant M13 clones were used as probes to isolate corresponding A phage clones, which could tentatively be mapped to the genome on the basis of presumed genetic collinearity between HHV-7 and HHV-6. Genomic 'walking' between EcoRI and BamHI 3, genomic libraries enabled overlapping neighbouring clones to be identified and mapped. Each of these clones was analysed to map BamHI, EcoRI, Sail, Smal and Xhol restriction endonuclease sites to provide complete endonuclease maps for the entire genome.

Original languageEnglish (US)
Pages (from-to)1901-1912
Number of pages12
JournalJournal of General Virology
Volume77
Issue number8
DOIs
StatePublished - Aug 1996

ASJC Scopus subject areas

  • Virology

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