Sensitive, high-resolution chromatin and chromosome mapping in situ

Presence and orientation of two closely integrated copies of EBV in a lymphoma line

Jeanne Bentley Lawrence, Carol A. Villnave, Robert H. Singer

Research output: Contribution to journalArticle

421 Citations (Scopus)

Abstract

Here we describe development and application of highly sensitive fluorescence methodology for localization of single-copy sequences in interphase nuclei and metaphase chromosomes by nonisotopic in situ hybridization. Application of this methodology to the investigation of Epstein-Barr virus integration in the Namalwa lymphoma cell line has revealed two EBV genomes closely integrated at the known site on chromosome 1. Detecting sequences as small as 5 kb, we further demonstrate resolution within interphase nuclei of two fragments of the viral genome spaced only 130 kb apart. Results indicate that the viral genomes are in opposite orientations and separated by roughly 340 kb of cellular DNA. This work demonstrates the feasibility and resolving power of interphase chromatin mapping to assess the proximity of closely spaced DNA sequences. Implications for virology, gene mapping, and investigation of nuclear organization are discussed.

Original languageEnglish (US)
Pages (from-to)51-61
Number of pages11
JournalCell
Volume52
Issue number1
DOIs
StatePublished - Jan 15 1988
Externally publishedYes

Fingerprint

Chromosome Mapping
Interphase
Chromosomes
Human Herpesvirus 4
Chromatin
Lymphoma
Genes
Viral Genome
Virus Integration
Virology
Chromosomes, Human, Pair 1
Metaphase
In Situ Hybridization
DNA sequences
Optical resolving power
Fluorescence
Viruses
Genome
Cell Line
DNA

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

Sensitive, high-resolution chromatin and chromosome mapping in situ : Presence and orientation of two closely integrated copies of EBV in a lymphoma line. / Bentley Lawrence, Jeanne; Villnave, Carol A.; Singer, Robert H.

In: Cell, Vol. 52, No. 1, 15.01.1988, p. 51-61.

Research output: Contribution to journalArticle

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