杂种细胞基因组文库的构建及单拷贝顺序的分离和顺序分析

THE ESTABLISHMENT OF A HYBRID GENOMIC LIBRARY AND ISOLATION AND SEQUENCE ANALYSIS OF UNIQUE SEQUENCE FROM SPECIFIC HUMAN CHROMOSOME

杂种细胞14-7-1中,含有完整的人体11号和20号染色体,同时还含有4号和8号的部分染色体。用体外包装的方法构建基因组文库。用种特异的探针分离出含有人体基因组顺序的重组子,分离得到8个单拷贝顺序。通过与已建立的杂种细胞克隆分布板杂交以及染色体的原位杂交方法。将1个单拷贝顺序FD11-1定位在11p11-q11上。这是1个767bp的核苷酸顺序,经1991年Genbank检索。没有找到任何具有同源性的基因或片段,这是1个新克隆的DNA片段;为临床诊断以及人类基因组全顺序测定提供有用的探针和座位。

Hybrid cell can 62 used as donor of human chromosomes. Recombinants containing human genome origin can be isolated from the hybrid cell genomic library by using speeies-specific probes. The unique sequences from specific human chromosome can be isolated from these recombinants by using proper restriction enzymes. A new FD11-1 DNA fragment of 767 bp was isolated from human chromosome 11 and analyzed.