Johnson-Pais TL, Leach RJ
Department of Cellular and Structural Biology, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, Texas, 78284
Methods. 1996 Feb;9(1):20-9. doi: 10.1006/meth.1996.0004.
Microcell hybrids are useful resources for the mapping of human chromosomes. The procedure of microcell-mediated chromosome transfer often causes fragmentation of the donor chromosome. These fragment-containing microcell hybrids frequently contain a limited region around the locus used in selecting for retention of the chromosome in the hybrids, as well as other fragments from the donor chromosome. Monochromosomal microcell hybrids are useful as the donor cell line for creation of radiation-reduced hybrids. In contrast to fragment-containing microcell hybrids, radiation-reduced hybrids can be used to construct maps of regions of chromosomes that lack selectable markers. For both fragment-containing hybrids and radiation-reduced hybrids, the presence or absence of chromosome-specific sequences can be determined and used to construct a linear map of the chromosome. A protocol and general overview outlining the key concepts in the construction and analysis of a radiation-reduced hybrid panel is presented.
微细胞杂种是人类染色体定位的有用资源。微细胞介导的染色体转移过程常常导致供体染色体的片段化。这些含有片段的微细胞杂种通常在用于选择杂种中染色体保留的位点周围含有有限区域,以及来自供体染色体的其他片段。单染色体微细胞杂种可用作创建辐射减少杂种的供体细胞系。与含有片段的微细胞杂种相反,辐射减少杂种可用于构建缺乏选择标记的染色体区域图谱。对于含有片段的杂种和辐射减少杂种,均可确定染色体特异性序列的存在与否,并用于构建染色体线性图谱。本文介绍了一个方案及总体概述,概述了辐射减少杂种细胞系构建和分析中的关键概念。