利用辐射杂种构建16号染色体图谱。
Construction of a map of chromosome 16 by using radiation hybrids.
作者信息
Ceccherini I, Romeo G, Lawrence S, Breuning M H, Harris P C, Himmelbauer H, Frischauf A M, Sutherland G R, Germino G G, Reeders S T
机构信息
Laboratorio di Genetica Molecolare, Istituto G. Gaslini, Genova, Italy.
出版信息
Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):104-8. doi: 10.1073/pnas.89.1.104.
A human-hamster cell hybrid carrying a single copy of chromosome 16 as the only human genetic material was irradiated with a single dose of gamma-rays (7000 rads; 1 rad = 0.01 Gy) and then fused with a thymidine kinase-deficient hamster cell line (RJKM) to generate radiation hybrids retaining unselected fragments of this human chromosome. In two experiments, 223 hybrids were isolated in hypoxanthine/aminopterine/thymidine (HAT) medium and screened with 38 DNA probes, corresponding to anonymous DNA or gene sequences localized on chromosome 16. The most likely order and location of the 38 DNA sequences were established by multiple pairwise analysis and scaled to estimate physical distance in megabases. The order and the distances thus obtained are mostly consistent with available data on genetic and physical mapping of these markers, illustrating the usefulness of radiation hybrids for mapping.
一个携带16号染色体单拷贝作为唯一人类遗传物质的人-仓鼠细胞杂种，用单剂量γ射线（7000拉德；1拉德 = 0.01戈瑞）进行照射，然后与胸腺嘧啶激酶缺陷的仓鼠细胞系（RJKM）融合，以产生保留该人类染色体未选择片段的辐射杂种。在两个实验中，在次黄嘌呤/氨基蝶呤/胸腺嘧啶核苷（HAT）培养基中分离出223个杂种，并用38个DNA探针进行筛选，这些探针对应于定位于16号染色体上的匿名DNA或基因序列。通过多重成对分析确定了38个DNA序列最可能的顺序和位置，并进行了缩放以估计以兆碱基为单位的物理距离。由此获得的顺序和距离大多与这些标记的遗传和物理图谱的现有数据一致，说明了辐射杂种在图谱绘制中的有用性。
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