Riethman H C, Moyzis R K, Meyne J, Burke D T, Olson M V
Department of Genetics, Washington University School of Medicine, Saint Louis, MO 63110.
Proc Natl Acad Sci U S A. 1989 Aug;86(16):6240-4. doi: 10.1073/pnas.86.16.6240.
Telomeric fragments of human DNA ranging in size from 50 to 250 kilobases were cloned into Saccharomyces cerevisiae using a yeast-artificial-chromosome (YAC) vector. Six human-telomeric YAC (HTY) strains were selected by virtue of the specific hybridization of their DNA with the human telomeric terminal-repeat sequence (TTAGGG)n, and the telomeric localization of this sequence within each YAC was demonstrated by its sensitivity to nuclease BAL-31. In situ hybridization of DNA from three of these HTY strains with human metaphase chromosomes yielded discrete patterns of hybridization signals at the telomeres of a limited number of human chromosomes, different for each clone. DNA from selected cosmid subclones of one of the HTY strains was used to localize the origin of the cloned telomeric DNA by in situ hybridization to the tip of the long arm of chromosome 7.
使用酵母人工染色体（YAC）载体，将大小在50至250千碱基之间的人类DNA端粒片段克隆到酿酒酵母中。通过其DNA与人端粒末端重复序列（TTAGGG）n的特异性杂交，筛选出6个人类端粒YAC（HTY）菌株，并且通过其对核酸酶BAL - 31的敏感性证明了该序列在每个YAC内的端粒定位。其中三个HTY菌株的DNA与人中期染色体的原位杂交在有限数量的人类染色体端粒处产生了离散的杂交信号模式，每个克隆的模式不同。使用来自其中一个HTY菌株的选定黏粒亚克隆的DNA，通过原位杂交到7号染色体长臂末端来定位克隆的端粒DNA的起源。
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