人21号染色体特异性黏粒的分离及其在染色体亚区域黏粒重叠群定位中的应用。

Isolation of human chromosome 21-specific cosmids and their uses in mapping of cosmid contigs on chromosomal subregions.

作者信息

Hou D X, Kishida H, Shimokawa T, Soeda E

机构信息

Gene Bank, Tsukuba Life Science Center, Institute of Physical and Chemical Research (RIKEN), Ibaraki, Japan.

出版信息

Jpn J Hum Genet. 1994 Dec;39(4):411-20. doi: 10.1007/BF01892386.

DOI:10.1007/BF01892386

PMID:7873753

Abstract

A cosmid library of 3 x 10(5) clones has been constructed from a human x hamster hybrid cell line, 153E9a3, which contains human chromosome 21 (HC21) as the only human chromosome. From 56,500 clones of this library, 229 HC21-specific cosmids have been isolated by their hybridization to total human DNA and by their failure to hybridize to total Chinese hamster DNA. The cosmids isolated were then characterized, of these, 28 cosmids (12.2% of those tested) contained Not1 site(s), and 41 cosmids were localized on the eight subregions of HC21 by differential hybridization with Alu-PCR products obtained from a hybrid mapping panel. The cosmids localized were further integrated into the existing contigs using the end-specific probes of the clone insert. Therefore, they provided useful anchor points for contig mapping and walking.

摘要

已从人 - 仓鼠杂交细胞系153E9a3构建了一个包含3×10⁵个克隆的黏粒文库，该细胞系仅含人类21号染色体（HC21）作为唯一的人类染色体。从该文库的56,500个克隆中，通过与总人类DNA杂交以及与总中国仓鼠DNA不杂交，分离出了229个HC21特异性黏粒。然后对分离出的黏粒进行了表征，其中28个黏粒（占测试黏粒的12.2%）含有Not1位点，通过与从杂交定位板获得的Alu - PCR产物进行差异杂交，41个黏粒定位在HC21的八个亚区域上。使用克隆插入片段的末端特异性探针将定位的黏粒进一步整合到现有的重叠群中。因此，它们为重叠群作图和步移提供了有用的锚定位点。