TY - JOUR
T1 - Sensitive, high-resolution chromatin and chromosome mapping in situ
T2 - Presence and orientation of two closely integrated copies of EBV in a lymphoma line
AU - Bentley Lawrence, Jeanne
AU - Villnave, Carol A.
AU - Singer, Robert H.
N1 - Funding Information:
We express our appreciation to David Ward for his suggestion that we Investigate the Namalwa line, and we thank Mark Heller and Eliott Kieff for useful discussions. Eliott Kieff provided the Namalwa cell line and Jim Skare provided clones of EBV fragments. We acknowledge the secretarial and administrative assistance of Elayn Byron, and the technical assistance of Lisa Marselle, Marie Georglo, and Lee Soeux. This work was supported by a Muscular Dystrophy grant to J. B. L. and National Institutes of Health grant HD18066 to R. S. and J B. L.
PY - 1988/1/15
Y1 - 1988/1/15
N2 - 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.
AB - 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.
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U2 - 10.1016/0092-8674(88)90530-2
DO - 10.1016/0092-8674(88)90530-2
M3 - Article
C2 - 2830981
AN - SCOPUS:0023867576
SN - 0092-8674
VL - 52
SP - 51
EP - 61
JO - Cell
JF - Cell
IS - 1
ER -