人类X染色体Xp11.3～21.3区部分YAC重叠群构建及大尺度物理图谱分析

Some YAC Contig Construction and Long Range Physical Mapping at Human X Chromosome Xp11.3～21.3

人类Ｘ染色体短臂１１．３～２１．３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重叠群构建及大尺度物理作图。用这一区域已知的探针（ＯＴＣ、２ｂＡ３、ＤＭＤｃＤＮＡ），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７７年阳性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指纹图谱分析和稀切点酶大尺度物理图谱分析。综合上述信息，对这些ＹＡＣ进行了排序，在Ｘｐ１１．３～２１．３区得到了６个ＹＡＣ重叠群，总共覆盖了约１５．３Ｍｂ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。

uman X chromosome short arm Xp113～p213 is an area,where several genetic disease gene loci are located.In this work,the YAC contig construction,long range physical mapping were done for this region.Some DNA probes and STS markers were used for YAC screening.Totally 77 YACs were obtained form the YAC libraries of CEPH,ICRF and ours.The size determination,26 pairs of microsatelite STS analysis,single copy probe hybridization,Alu-PCR finger printing and long range physical mapping were conducted with these YACs.These results allowed us to map these YACs,and finally 6 YAC contigs were obtained in Xp113～213,covering about 153 Mb.This work will greatly facilitate the positional cloning of disrase genes or the genome sequencing in this important region.