Wang C Y, Shi J D, Huang Y Q, Cruz P E, Ochoa B, Hawkins-Lee B, Davoodi-Semiromi A, She J X
Immunology and Laboratory Medicine, University of Florida, Gainesville, Florida, 32610, USA.
Genomics. 1999 Aug 15;60(1):12-9. doi: 10.1006/geno.1999.5908.
Urofacial (Ochoa) syndrome is an autosomal recessive disease characterized by distorted facial expression and urinary abnormalities. Previously, we mapped the UFS gene to chromosome 10q23-q24 and narrowed the interval to one YAC clone of 1410 kb. Here, we have constructed a BAC/PAC contig of the 1-Mb region using STS content mapping with 42 BAC/PAC-end sequences, 9 previously reported and 16 newly identified microsatellite markers, and 14 EST markers. A total of 26 polymorphic microsatellite markers were genotyped for 31 UFS patients from Colombia and 2 patients from the United States. Haplotype analyses suggest that the UFS gene is located within two overlapping BAC clones, a region of <360 kb of DNA sequence. We tested 42 EST markers previously mapped to the D10S1709-D10S603 interval against the BAC/PAC contig and identified 11 ESTs located in the 1-Mb region. Four of the 11 ESTs mapped to the 360-kb UFS critical region. Shotgun sequencing of the two BAC clones and BLASTN search of the EST databases revealed 3 other ESTs contained in the UFS critical region. These results will facilitate the cloning and identification of the UFS gene.
泌尿生殖面部（奥乔亚）综合征是一种常染色体隐性疾病，其特征为面部表情扭曲和泌尿异常。此前，我们已将泌尿生殖面部综合征基因定位到10号染色体的10q23 - q24区域，并将区间缩小至一个1410 kb的酵母人工染色体（YAC）克隆。在此，我们利用STS含量作图法构建了一个1兆碱基（Mb）区域的细菌人工染色体（BAC）/ 噬菌体P1人工染色体（PAC）重叠群，该方法使用了42个BAC / PAC末端序列、9个先前报道的微卫星标记、16个新鉴定的微卫星标记以及14个表达序列标签（EST）标记。对来自哥伦比亚的31名泌尿生殖面部综合征患者和来自美国的2名患者的26个多态性微卫星标记进行了基因分型。单倍型分析表明，泌尿生殖面部综合征基因位于两个重叠的BAC克隆内，即一个小于360 kb的DNA序列区域。我们针对BAC / PAC重叠群对先前定位到D10S1709 - D10S603区间的42个EST标记进行了检测，并在1 Mb区域内鉴定出11个EST。这11个EST中的4个定位到了360 kb的泌尿生殖面部综合征关键区域。对这两个BAC克隆进行鸟枪法测序，并在EST数据库中进行BLASTN搜索，结果显示泌尿生殖面部综合征关键区域还包含另外3个EST。这些结果将有助于泌尿生殖面部综合征基因的克隆与鉴定。
