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首页> 外文期刊>Molecular and Cellular Biology >Construction of microcell hybrid clones containing specific mouse chromosomes: application to autosomes 8 and 17.
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Construction of microcell hybrid clones containing specific mouse chromosomes: application to autosomes 8 and 17.

机译：包含特定小鼠染色体的微细胞杂种克隆的构建：应用于常染色体8和17。

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Fibroblast cultures prepared from mice homozygous for a Robertsonian translocation (centric fusion) between autosomes 8 and 17 [Rb(8.17)] were used as donors in microcell-mediated chromosome transfer experiments. By using hamster recipient cells deficient in adenine phosphoribosyltransferase (APRT-) and selecting for expression of murine APRT (a chromosome 8 marker), microcell hybrids were isolated which retained only the mouse Rb(8.17) translocation in addition to the hamster chromosome complement. The translocation was stable in cells maintained under APRT+ selective pressure, and mouse marker traits encoded by genes on both chromosomes 8 and 17 segregated concordantly. A second family of hybrid clones was constructed by fusing microcells derived from wild-type mouse fibroblasts with APRT- hamster cells. Four of six clones analyzed retained only mouse chromosome 8. These studies demonstrated that microcell hybrids containing specific Robertsonian translocations as the only donor-derived genetic material can be obtained. Furthermore, a number of Robertsonian translocations between chromosomes which carry selectable markers (chromosomes 3, 8, and 11) and other autosomes have been described. By using fibroblast cultures prepared from mice containing these translocations as donors in microcell fusions, 18 of the 20 mouse chromosomes could be selectively fixed in different hybrid clones. Thus, a collection of 20 hybrid clones, each containing a single, specific mouse chromosome, can be constructed by using the strategy described in this report. The potential utility of such a monochromosomal hybrid panel is discussed.
机译：由小鼠纯合体制备的成纤维细胞培养物用于常染色体8和17 [Rb（8.17）]之间的Robertsonian易位（中心融合），用作微细胞介导的染色体转移实验的供体。通过使用缺乏腺嘌呤磷酸核糖基转移酶（APRT-）的仓鼠受体细胞并选择鼠APRT（8号染色体标记）的表达，分离了微细胞杂种，除仓鼠染色体补体外，其仅保留了小鼠Rb（8.17）易位。在APRT +选择性压力下维持的细胞中，转位稳定，并且由8号和17号染色​​体上的基因编码的小鼠标记性状一致地分离。通过将衍生自野生型小鼠成纤维细胞的微细胞与APRT-仓鼠细胞融合，构建了第二个杂种克隆家族。分析的六个克隆中有四个仅保留了小鼠8号染色体。这些研究表明，可以获得包含特定罗伯逊易位作为唯一的供体来源遗传材料的微细胞杂种。此外，已经描述了在带有选择标记（染色体3、8和11）和其他常染色体的染色体之间的许多罗伯逊易位。通过使用由包含这些易位的小鼠制备的成纤维细胞培养物作为微细胞融合中的供体，可以选择性地将20条小鼠染色体中的18条固定在不同的杂种克隆中。因此，可以使用本报告中描述的策略构建20个杂种克隆的集合，每个克隆包含一个特定的小鼠染色体。讨论了这种单色杂交板的潜在用途。

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