本文選題：GJB2基因　切入點：12SrRNA基因突變　出處：《南京醫(yī)科大學(xué)》2013年碩士論文

【摘要】：第一部分：非綜合征型耳聾患者GJB2基因及線粒體DNA12S rRNA基因突變分析 目的：通過對非綜合征型耳聾患者進行GJB2基因及線粒體DNA12SrRNA基因的突變檢測，了解其突變類型和突變頻率，為進一步指導(dǎo)聾病基因篩查和遺傳咨詢提供理論依據(jù)。 方法：收集190例門診非綜合征耳聾患者的外周血樣，常規(guī)法提取基因組DNA。PCR擴增GJB2基因編碼區(qū)及線粒體DNA12S rRNA目的片段，對純化后的目的片段進行測序分析，識別耳聾相關(guān)的致病突變。 結(jié)果：190例患者中，，128例攜帶GJB2突變，共23種突變類型，其中已知的致聾突變5種，以235delC為主（16.32%）；發(fā)現(xiàn)42個線粒體DNA12SrRNA突變位點，43種突變類型，其中明確與非綜合征型耳聾相關(guān)的致病突變3種：C1494T, A1555G和961位點突變，以A1555G為主（6.32%）。檢測出GJB2單雜合致聾突變攜帶率者25例（13.16%），其中235delC單雜合突變攜帶者14例（7.37%）。5名患者同時攜帶GJB2235delC單雜合突變和線粒體DNA12SrRNA突變。 結(jié)論：GJB2基因和線粒體DNA12S rRNA基因是我國非綜合征型耳聾患者的突變熱點基因，針對上述基因的突變篩查有助于明確耳聾的分子病因。鑒于GJB2235delC單雜合突變攜帶率較高，進一步研究其致聾機制是必要的。 第二部分：GJB2235delC單雜合突變耳聾患者的GJB2基因上游調(diào)控區(qū)序列分析 目的：探討GJB2基因上游調(diào)控區(qū)的序列變化與該基因編碼區(qū)235delC單雜合突變致聾的相關(guān)性。 方法：對實驗室前期確定的14例攜帶GJB2235delC單雜合突變的耳聾患者、20例未發(fā)現(xiàn)致病突變的耳聾患者及25例聽力正常個體進行GJB2基因上游調(diào)控區(qū)序列的PCR擴增及測序分析。 結(jié)果：在14例GJB2235delC雜合突變耳聾患者中檢測到兩種堿基變異-843AC和-195TC，結(jié)合對照組檢測結(jié)果，基本可確定兩者均為多態(tài)性改變。 結(jié)論：GJB2基因上游調(diào)控區(qū)序列的改變可能不是協(xié)同該基因編碼區(qū)突變致聾的常見分子事件。
[Abstract]:Part I: mutation analysis of GJB2 gene and mitochondrial DNA12S rRNA gene in patients with non-syndromic deafness. Objective: to investigate the mutation types and frequencies of GJB2 gene and mitochondrial DNA12SrRNA gene in patients with non-syndromic deafness, and to provide theoretical basis for gene screening and genetic counseling of deafness. Methods: peripheral blood samples were collected from 190 patients with non-syndromic deafness. Genomic DNA.PCR was extracted to amplify the coding region of GJB2 gene and the target fragment of mitochondrial DNA12S rRNA, and the purified fragment was sequenced. Identify deaf-related mutations. Results among the 190 patients, 128 had GJB2 mutations, including 23 mutation types, 5 of which were known deafness mutations, mainly 235delC 16.322.43 mutations were found in 42 mitochondrial DNA12SrRNA mutation loci. Among them, there were 3 kinds of mutations at the 1: C1494T, A1555G and 961 loci that were specifically associated with non-syndromic deafness. 25 cases with GJB2 single heterozygous deafness mutation were detected, among which 14 patients with 235delC single heterozygous mutation carried GJB2235delC monozygosity mutation and mitochondrial DNA12SrRNA mutation at the same time. Conclusion the gene of: GJB2 and mitochondrial DNA12S rRNA are the hot mutation genes in non-syndromic deafness patients in China. The mutation screening of these genes is helpful to clarify the molecular etiology of deafness. In view of the high rate of single heterozygosity of GJB2235delC, It is necessary to further study the mechanism of deafness. Part two: sequence analysis of upstream regulatory region of GJB2 gene in deafness patients with single heterozygous mutation GJB2235delC. Aim: to investigate the relationship between the sequence change of upstream regulatory region of GJB2 gene and the deafness caused by 235delC single heterozygosity mutation in the coding region of GJB2 gene. Methods: PCR amplification and sequencing analysis of the upstream regulatory region of GJB2 gene were performed in 14 deafness patients with single GJB2235delC heterozygosity and 25 hearing normal individuals. Results: two kinds of base mutation -843AC and -195TC were detected in 14 patients with GJB2235delC heterozygous deafness. Conclusion the change of the upstream regulatory region of the gene may not be a common molecular event of deafness caused by the mutation in the coding region of the gene.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R764.43

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