两个氨基糖甙类药物性耳聋家系的分子遗传学研究
发布时间：2018-09-18 16:03
【摘要】：目的: 探讨2个氨基糖甙类药物所致非综合征型耳聋中国家系的分子遗传学特征。 方法: 从2个氨基糖甙类药物性耳聋及非综合征型耳聋家系选择部分成员,包括7名母系成员及1名听力正常配偶,取外周血,提取基因组DNA。用遗传性耳聋基因检测芯片检测4个中国人中常见的耳聋相关基因中的9个热点突变,包括GJB2 ( 35delG,176del16bp,235delC及299delAT ),GJB3 ( C538T ),SLC26A4( IVS7-2AG,A2168G )和线粒体DNA 12S rRNA ( A1555G,C1494T )。PCR扩增阳性突变基因并测序验证检测结果。然后,采用特定引物对两家系的先证者分别进行线粒体DNA全序列及核基因TRMU和MTO1编码区的PCR扩增,扩增产物经纯化后直接用ABI 3730型测序仪测序,并将测序结果与标准序列比对,识别有意义的碱基变异。 结果: 芯片检测发现,两家系的7名母系成员均存在同质性线粒体DNA12S rRNA基因C1494T突变。与修正的剑桥参考序列相比,两名先证者的线粒体DNA全序列分析共发现了53个碱基变异,但除已知的12S rRNA C1494T突变外,其余52个碱基变异均为已报道的多态性位点;两家系先证者线粒体单体型分别是D4和D5a。TRMU和MTO1基因序列分析无异常发现。 结论: 线粒体DNA 12S rRNA C1494T突变是这两个家系耳聋发生的主要分子基础,而氨基糖甙类抗生素的应用增强了该突变的表型表达;未能证实线粒体单体型以及核基因TRMU和MTO1对家系成员C1494T突变的表型具有修饰作用。
[Abstract]:Objective: to investigate the molecular genetic characteristics of two national strains of non-syndromic deafness induced by aminoglycosides. Methods: genomic DNA. was extracted from peripheral blood from two families of aminoglycoside drug induced deafness and non-syndromic deafness, including 7 matrilineal members and 1 normal hearing spouse. Genetic deafness gene detection microarray was used to detect 9 hot spot mutations of deaf-related genes in 4 Chinese. GJB3 (C538T), SLC26A4 (IVS7-2AG,A2168G) and mitochondrial DNA 12S rRNA (A1555GfC1494T) were amplified by GJB2 (35delGf176del16bpc235delC and 299delAT), and the results were verified by sequencing. Then, specific primers were used to amplify the whole mitochondrial DNA sequence and the nucleotide TRMU and MTO1 coding region of the two families, respectively. After purification, the amplified products were sequenced directly by ABI 3730 type sequencer, and the sequencing results were compared with the standard sequence. Identify meaningful base mutations. Results: the results of microarray analysis showed that the homogenous mitochondrial DNA12S rRNA gene C1494T mutation was found in 7 maternal members of both families. Compared with the modified Cambridge reference sequence, 53 nucleotide mutations were found in mitochondrial DNA of the two proband, except for the 12s rRNA C1494T mutation, the other 52 nucleotide mutations were reported polymorphism sites. The mitochondrial haplotypes of two families were D4, D5a.TRMU and MTO1, respectively. Conclusion: mitochondrial DNA 12s rRNA C1494T mutation is the main molecular basis of deafness in these two families, and the application of aminoglycoside antibiotics enhances the expression of the mutant phenotype. It was not confirmed that mitochondrial haplotype and nuclear gene TRMU and MTO1 could modify the phenotype of C1494T mutation in family members.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R764
本文编号：2248415
[Abstract]:Objective: to investigate the molecular genetic characteristics of two national strains of non-syndromic deafness induced by aminoglycosides. Methods: genomic DNA. was extracted from peripheral blood from two families of aminoglycoside drug induced deafness and non-syndromic deafness, including 7 matrilineal members and 1 normal hearing spouse. Genetic deafness gene detection microarray was used to detect 9 hot spot mutations of deaf-related genes in 4 Chinese. GJB3 (C538T), SLC26A4 (IVS7-2AG,A2168G) and mitochondrial DNA 12S rRNA (A1555GfC1494T) were amplified by GJB2 (35delGf176del16bpc235delC and 299delAT), and the results were verified by sequencing. Then, specific primers were used to amplify the whole mitochondrial DNA sequence and the nucleotide TRMU and MTO1 coding region of the two families, respectively. After purification, the amplified products were sequenced directly by ABI 3730 type sequencer, and the sequencing results were compared with the standard sequence. Identify meaningful base mutations. Results: the results of microarray analysis showed that the homogenous mitochondrial DNA12S rRNA gene C1494T mutation was found in 7 maternal members of both families. Compared with the modified Cambridge reference sequence, 53 nucleotide mutations were found in mitochondrial DNA of the two proband, except for the 12s rRNA C1494T mutation, the other 52 nucleotide mutations were reported polymorphism sites. The mitochondrial haplotypes of two families were D4, D5a.TRMU and MTO1, respectively. Conclusion: mitochondrial DNA 12s rRNA C1494T mutation is the main molecular basis of deafness in these two families, and the application of aminoglycoside antibiotics enhances the expression of the mutant phenotype. It was not confirmed that mitochondrial haplotype and nuclear gene TRMU and MTO1 could modify the phenotype of C1494T mutation in family members.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R764
【参考文献】
相关期刊论文 前1条
1 ;Mitochondrial rRNA and tRNA and hearing function[J];Cell Research;2007年03期
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