采用基于序列标签位点（STS）的方法在酵母人工染色体中构建跨越人类肌营养不良蛋白基因的2.6兆碱基重叠群。

Construction of a 2.6-Mb contig in yeast artificial chromosomes spanning the human dystrophin gene using an STS-based approach.

作者信息

Coffey A J, Roberts R G, Green E D, Cole C G, Butler R, Anand R, Giannelli F, Bentley D R

机构信息

Paediatric Research Unit, United Medical School of Guy's Hospital, London, United Kingdom.

出版信息

Genomics. 1992 Mar;12(3):474-84. doi: 10.1016/0888-7543(92)90437-w.

DOI:10.1016/0888-7543(92)90437-w

PMID:1559699

Abstract

A sequence tagged site (STS)-based approach has been used to construct a 2.6-Mb contig in yeast artificial chromosomes (YACs) spanning the human dystrophin gene. Twenty-seven STSs were used to identify and overlap 34 YAC clones. A DNA fingerprint of each clone produced by direct Alu-PCR amplification of YAC colonies and the isolation of YAC insert ends by vectorette PCR were used to detect overlaps in intron 1 (280 kb) where no DNA sequence data were available, thereby achieving closure of the map. This study has evaluated methods for mapping large regions of the X chromosome and provides a valuable resource of the dystrophin gene in cloned form for detailed analysis of gene structure and function in the future.

摘要

一种基于序列标签位点（STS）的方法已被用于在酵母人工染色体（YAC）中构建一个跨越人类肌营养不良蛋白基因的2.6兆碱基重叠群。使用27个STS来鉴定并重叠34个YAC克隆。通过对YAC菌落进行直接Alu-PCR扩增产生每个克隆的DNA指纹图谱，并通过载体PCR分离YAC插入末端，用于检测内含子1（280 kb）中没有DNA序列数据的重叠区域，从而完成图谱的封闭。这项研究评估了绘制X染色体大片段区域的方法，并为未来详细分析基因结构和功能提供了克隆形式的肌营养不良蛋白基因的宝贵资源。