Four novel and two previously reported mutations of the PAX6 gene in patients with aniridia.
Four novel and two previously reported mutations of the PAX6 gene in patients with aniridia.
复制标题
无虹膜患者中 PAX6 基因的四种新突变和两种先前报道的突变。
DOI：
10.1002/humu.1380110167
复制
发表时间：
1998
期刊：
影响因子：
3.9
通讯作者：
Chao,LY
中科院分区：
文献类型：
--
作者：
Saunders,GF;Chao,LY
无虹膜是一种先天性双侧眼病，其特征是虹膜完全或部分缺失和虹膜发育不良。无虹膜基因定位于1号染色体lp13(Compton等人，1988；Davis等人，1989；Gessler等人，1989；Rose等人，1990；Lyons等人，1992)，是通过定位克隆(ton等人，1991)分离出来的，与啮齿动物(Hill等人，1991；Walther和Gruss，1991；ton等人，1992；Matsuo等人，1993)和许多其他动物的Tie Pax6基因高度同源。这些基因包含两个高度保守的基序，即配对的框和同源框区域。人类PAX6基因全长22kb，由12个内含子和13个外显子组成(格拉泽等人，1992)。PAX6基因的突变导致啮齿类动物的小眼睛表型(Hill等人，1991)和人的无虹膜表型(Gaser等人，1992；Jordan等人，1992)。在这项研究中，我们用聚合酶链式反应、异源双链分析和测序的方法检测了6例无虹膜患者的PAX6基因。用盐析法从血液中提取宪法DNA(Miller等人，1988年)。我们用来扩增PAX6基因外显子的聚合酶链式反应引物是格拉泽等人(1992)的。聚合酶链式反应在Perkin-Elmer的聚合酶链式反应缓冲液中进行，反应缓冲液中含有放射性的[01-32P]dCTP和每对引物所特有的氯化镁浓度。聚合酶链式反应参数为：97“C/4min，然后在94”、60“和72℃下分别进行35个循环，50s、1min和40s。
Aniridia is a congenital, bilateral ocular disorder characterized by the complete or partial absence of the iris and iris hypoplasia. The aniridia gene, located on chromosome 1 lp13 (Compton et al., 1988; Davis et al., 1989; Gessler et al., 1989; Rose et al., 1990; Lyons et al., 1992), was isolated by positional cloning (Ton et al., 1991) and is highly homologous with tie Pax6 gene of rodents (Hill et al., 1991; Walther and Gruss, 1991; Ton et al., 1992; Matsuo et al., 1993) and many other animals.PAX6 belongs to the paired-like class of developmental genes first described in Drosophila. These genes contain two highly conserved motifs, the paired box and homeobox regions. The human PAX6 gene spans 22 kb and contains 12 introns flanked by 13 exons (Glaser et al., 1992). Mutations in the PAX6 gene result in the small-eye phenotype in rodents (Hill et al., 1991) and the aniridia phenotype in hu-mans (Glaser et al., 1992; Jordan et al., 1992). In this study, we examined the PAX6 gene in six aniridia patients by polymerase chain reaction, heteroduplex analysis, and sequencing. Constitutional DNA was extracted from blood by using the saltingout method (Miller et al., 1988). The PCR primers we used for the exons of PAX6 were those of Glaser et al.(1992). PCR reactions were performed in Perkin-Elmer's PCR reaction buffer with radioactive [01-32P] dCTP and an MgClz concentration specific for each primer pair. The PCR parameters were: 97" C/4 min, followed by 35 cycles of 50 sec at 94", 1 min at 60", and 40 sec at 72 C.