权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Diagnosis of xeroderma pigmentosum patients and carriers by PCR-RFLP analysis

PCR-RFLP分析对着色性干皮病患者和携带者的诊断

基本信息

批准号：
06670864
负责人：
UEDA Masato
金额：
$ 1.41万
依托单位：
Kobe University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06670864/
关键词：
xeroderma pigmentosum XPA gene PCR-RFLP gene diagnosis amniotic fluid p53 PCR-RFLP法 XPAC遺伝子 PCR‐RFLP法

项目摘要

The gene responsible for xeroderma pigmentosum (XP) group A has recently been cloned and designated XPA gene. Previous studies have shown that most Japanese XPA patients have homozygous mutations for the splicing site of intron 3 of the XPA gene, which was recognized by restriction endonuculease (RE) AlwNI (AlwNI mutation). Other mutations found to data have been the nonsense mutation at codon 228 in exon 6, recognized by RE HphI (HphI mutation), and at codon 116 in exon 3, recognized by RE MseI (MseI mutation) . Using polymerase chain reaction restriction fragment length polymorphism (PCR-PFLP) analysis, we examined the point mutations of the XPA gene.(1) We found that 12 patients of 17 patients were homozygous for the AlwNI mutation, four were compound heterozygotes for the AlwNI mutation and the HphI mutation, and one was a compound heterozygote for the AlwNI mutation and the MseI mutation. Investigation of their clinical features suggested that the four patients with HphI mutation … More had milder clinical manifestations. PCR-RFLP analysis of the XPA gene in the three asymptomatic siblings of the XPA patients revealed that two were carriers of the mutated XPA allele, and one was not a carrier.(2) We found two siblings with XP group A who showed a significant difference in clinical manifestations and younger sister was much milder. The elder sister started strict sun protection at 4 years of age, whereas the younger began at 2 years of age. PCR-RFLP analysis revealed that both patients had the identical mutation in XPA gane. These data suggent, that in patients with XP the earlier sun protection begins had the later skin cancer develops.(3) We diagnosed the XPA gene mutation of a fetus whose sibling is a XPA patient. PCR-RFLP analysis of DNA from amniotic fluid showed the homozygous mutation.(4) P53 protein is critical for DNA damage repair, cell cycle and apoptosis. The p53 protein induction by UVB in XPA cells with various gene mutations was examined using western blot analysis. Cells with HphI/AlwNI heterozygous mutations showed weaker induction of p53 protein by UVB than cells with AlwNI homozygous mutations or AlwNI/MseI heterozygous mutations. Less

色素性干皮病(XP)A组致病基因最近被克隆并命名为XPA基因。以往的研究表明，大多数日本XPA患者存在XPA基因内含子3剪接位点的纯合子突变，这被限制性内切酶(RE)AlwNI(AlwNI突变)识别。在数据中发现的其他突变是由RE HphI识别的第6外显子第228位密码子的无义突变(HphI突变)和由RE MseI识别的第3外显子第116位密码子的无义突变(MseI突变)。采用聚合酶链式反应-限制性片段长度多态性分析方法检测XPA基因点突变：(1)17例患者中有12例为AlwNI突变纯合子，4例为AlwNI突变和HphI突变复合杂合子，1例为AlwNI突变和MseI突变复合杂合子。对他们的临床特征的研究表明，四名携带HphI突变…的患者临床表现较轻者较多。对XPA患者的三个无症状同胞进行XPA基因的聚合酶链式反应-限制性片段长度多态性分析，发现两个XPA突变等位基因携带者，一个不是携带者。(2)我们发现两个XPA患者同胞的临床表现有显著差异，妹妹病情较轻。姐姐从4岁开始严格防晒，而妹妹从2岁开始防晒。聚合酶链式反应-限制性片段长度多态性分析显示，两例患者均存在XPA基因相同突变。这些数据表明，在XP患者中，开始防晒的时间越早，皮肤癌的发生就越晚。(3)我们诊断了一个胎儿的XPA基因突变，该胎儿的兄弟姐妹是XPA患者。对羊水DNA进行PCR-RFLP分析，证实为纯合子突变。(4)P53蛋白在DNA损伤修复、细胞周期和细胞凋亡中起关键作用。用免疫印迹法检测UVB对不同基因突变的XPA细胞P53蛋白的诱导作用。HphI/AlwNI杂合突变细胞对P53蛋白的诱导作用弱于AlwNI纯合子突变细胞和AlwNI/MseI杂合突变细胞。较少