Lawrence J B, Villnave C A, Singer R H
Department of Anatomy, University of Massachusetts Medical School, Worcester 01605.
Cell. 1988 Jan 15;52(1):51-61. doi: 10.1016/0092-8674(88)90530-2.
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.
在此，我们描述了一种高灵敏度荧光方法的开发与应用，该方法通过非同位素原位杂交来定位间期细胞核和中期染色体中的单拷贝序列。将此方法应用于研究爱泼斯坦 - 巴尔病毒（Epstein - Barr virus，EBV）在Namalwa淋巴瘤细胞系中的整合情况，结果显示有两个EBV基因组紧密整合在1号染色体的已知位点上。我们能够检测低至5 kb的序列，进一步证明了在间期细胞核中可分辨病毒基因组的两个片段，它们仅相隔130 kb。结果表明，病毒基因组呈相反方向，且被大约340 kb的细胞DNA隔开。这项工作证明了间期染色质图谱分析在评估紧密间隔的DNA序列接近程度方面的可行性和分辨率。文中还讨论了其对病毒学、基因图谱绘制以及细胞核组织研究的意义。
