通过对人类染色体区域11q23进行显微切割并分离酵母人工染色体所获得的微克隆的序列标签位点

Sequence tagged sites of microclones obtained by microdissection of a human chromosomal region 11q23 and isolation of yeast artificial chromosomes.

作者信息

Seki N, Yamauchi M, Saito T, Harada Y, Hori T

机构信息

Division of Genetics, National Institute of Radiological Sciences, Chiba, Japan.

出版信息

Jpn J Hum Genet. 1994 Jun;39(2):249-54. doi: 10.1007/BF01876845.

DOI:10.1007/BF01876845

PMID:8086642

Abstract

A human chromosomal region 11q23-specific DNA library has been constructed by means of microdissection-microcloning method with polymerase chain reaction (PCR) technique (Seki et al., Genomics 16: 1993). DNA sequences were determined for 25 microclones that contained approximately 300-500 bp insert and gave a unique (single copy) signal in Southern blot analysis. The sequence tagged site (STS) was designed and appropriate condition for PCR was determined for each unique microclone. Twelve STSs were established and used for PCR-screening of human genomic libraries constructed with yeast artificial chromosome (YAC). Thirteen YAC clones have been isolated from eight STSs. These chromosomal region-specific STSs and YAC clones will be useful in the positional cloning of disease-related genes localized to the q23 region of chromosome 11.

摘要

利用显微切割-微克隆方法和聚合酶链反应（PCR）技术构建了一个人类染色体区域11q23特异性DNA文库（关等，《基因组学》16：1993）。测定了25个微克隆的DNA序列，这些微克隆含有约300-500bp的插入片段，并且在Southern印迹分析中给出独特（单拷贝）信号。为每个独特的微克隆设计了序列标签位点（STS）并确定了PCR的合适条件。建立了12个STS并用于对用酵母人工染色体（YAC）构建的人类基因组文库进行PCR筛选。已从8个STS中分离出13个YAC克隆。这些染色体区域特异性STS和YAC克隆将有助于定位克隆定位于11号染色体q23区域的疾病相关基因。