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人类10号染色体的图谱整合:基于可靠遗传图谱的染色体特异性体细胞杂种细胞系和基因组克隆的分子与细胞遗传学分析

Map integration at human chromosome 10: molecular and cytogenetic analysis of a chromosome-specific somatic cell hybrid panel and genomic clones, based on a well-supported genetic map.

作者信息

Marzella R, Kokkinaki M A, Kapsetaki M, Ricco A, Argyrokastritis A, Kamakari S, Sarafidou T, Archidiacono N, Roussou A, Pasparaki A, Rocchi M, Moschonas N K

机构信息

Istituto di Genetica, Bari, Italy.

出版信息

Cytogenet Cell Genet. 1997;79(3-4):257-65. doi: 10.1159/000134738.

Abstract

Well-characterized, chromosome-specific somatic cell hybrid panels are powerful tools for the analysis of the human genome. We have characterized a panel of human x hamster somatic cell hybrids retaining fragments of human chromosome 10 by fluorescence in situ hybridization and associated them to genetic markers. Most of the hybrids were generated by the radiation-reduction method, starting from a chromosome 10-specific monochromosomal hybrid, whereas some were collected from hybrids retaining chromosome 10-specific fragments as a result of spontaneous in vitro rearrangements. PCR was used to score the retention of 57 microsatellite markers evenly distributed along a well-supported framework genetic map containing 149 loci uniquely placed at 69 anchor points (odds exceeding 1,000:1), with an average spacing of 2.8 cM. As an additional resource for genomic studies involving human chromosome 10, we report the cytogenetic localization of a series of YAC and PAC clones recognized by at least one genetic marker. Somatic cell hybrids provide a powerful source of partial chromosome paints useful for detailed clinical cytogenetic and primate chromosome evolution investigations. Furthermore, correlation of the above physical, genetic, and cytogenetic data contribute to an emerging consensus map of human chromosome 10.

摘要

特征明确的、染色体特异性体细胞杂种面板是分析人类基因组的有力工具。我们通过荧光原位杂交对一组保留人类10号染色体片段的人×仓鼠体细胞杂种进行了特征分析,并将它们与遗传标记相关联。大多数杂种是通过辐射减少法从一个10号染色体特异性单染色体杂种开始产生的,而有些则是从因体外自发重排而保留10号染色体特异性片段的杂种中收集的。PCR用于对57个微卫星标记的保留情况进行评分,这些标记沿着一个包含149个位点、唯一位于69个锚定点(优势比超过1000:1)的可靠框架遗传图谱均匀分布,平均间距为2.8厘摩。作为涉及人类10号染色体的基因组研究的额外资源,我们报告了一系列至少被一个遗传标记识别的YAC和PAC克隆的细胞遗传学定位。体细胞杂种为部分染色体涂染提供了强大的来源,可用于详细的临床细胞遗传学和灵长类染色体进化研究。此外,上述物理、遗传和细胞遗传学数据的相关性有助于形成一个新兴的人类10号染色体共识图谱。

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