人类X染色体Xp21．3－11．3区YAC重叠群的构建

The YAC Contig Construction for Human X Chromosome Xp21. 3-11. 3

人类Ｘ染色体短臂２１．３—１１．３区是含有视网膜色素变性等数种遗传病基因位点的区域。我们对这个具有重要医学生物学意义的区段进行了ＹＡＣ重叠群构建。用这一区域已知的探针（ＯＴＣ、ＤＸＳ１６６、ＤＭＤｃＤＮＡ）及ＳＴＳ位标，以ＹＡＣ菌落原位杂交法及ＰＣＲ法进行了ＹＡＣ的筛选；也采用了法国ＣＥＰＨ和英国ＩＣＲＦ的部分ＹＡＣ；总共得到了５５个阳性ＹＡＣ。对上述ＹＡＣ进行了长度测定，２６对微卫星ＳＴＳ图谱分析，单拷贝探针的杂交定位，Ａｌｕ－ＰＣＲ指纹图谱分析。综合上述信息，对这些ＹＡＣ进行了排序，在Ｘｐ２１．３－１１．３区得到了６个０ＹＡＣ重叠群，覆盖了约１５Ｍｂ的范围。这些ＹＡＣ重叠群的构建为开展该区域疾病基因的定位克隆（Ｐｏｓｉｔｉｏｎａｌｃｌｏｎｉｎｇ）及ＤＮＡ顺序测定奠定了基础。

Human X chromosome short arm Xp21. 3-p11. 3 is an area which contains several genetic disease gene lo'ci. In this work, the YAC contig construction was done for this region. Some DNA probes and STS markers was used for YAC screening. Totally 55 YACs were obtained from the YAC libraries of CEPH,ICRF and ours.The size determination, 26 pairs of microsatelite STS analysis, single copy probe hybridization and Alu-PCR fingerprinting were Performed with these YACs. These results allowed us to map these YACs, and finally 6 YAC contigs were obtained in Xp21. 3-11.3,covering about 15 Mb.This work will greatly facilitate the positional cloning of disease genes or the genome sequencins in this important region.