兩個(gè)氨基糖甙類藥物性耳聾家系的分子遺傳學(xué)研究
發(fā)布時(shí)間：2018-09-18 16:03
【摘要】：目的: 探討2個(gè)氨基糖甙類藥物所致非綜合征型耳聾中國家系的分子遺傳學(xué)特征。 方法: 從2個(gè)氨基糖甙類藥物性耳聾及非綜合征型耳聾家系選擇部分成員,包括7名母系成員及1名聽力正常配偶,取外周血,提取基因組DNA。用遺傳性耳聾基因檢測芯片檢測4個(gè)中國人中常見的耳聾相關(guān)基因中的9個(gè)熱點(diǎn)突變,包括GJB2 ( 35delG,176del16bp,235delC及299delAT ),GJB3 ( C538T ),SLC26A4( IVS7-2AG,A2168G )和線粒體DNA 12S rRNA ( A1555G,C1494T )。PCR擴(kuò)增陽性突變基因并測序驗(yàn)證檢測結(jié)果。然后,采用特定引物對兩家系的先證者分別進(jìn)行線粒體DNA全序列及核基因TRMU和MTO1編碼區(qū)的PCR擴(kuò)增,擴(kuò)增產(chǎn)物經(jīng)純化后直接用ABI 3730型測序儀測序,并將測序結(jié)果與標(biāo)準(zhǔn)序列比對,識別有意義的堿基變異。 結(jié)果: 芯片檢測發(fā)現(xiàn),兩家系的7名母系成員均存在同質(zhì)性線粒體DNA12S rRNA基因C1494T突變。與修正的劍橋參考序列相比,兩名先證者的線粒體DNA全序列分析共發(fā)現(xiàn)了53個(gè)堿基變異,但除已知的12S rRNA C1494T突變外,其余52個(gè)堿基變異均為已報(bào)道的多態(tài)性位點(diǎn);兩家系先證者線粒體單體型分別是D4和D5a。TRMU和MTO1基因序列分析無異常發(fā)現(xiàn)。 結(jié)論: 線粒體DNA 12S rRNA C1494T突變是這兩個(gè)家系耳聾發(fā)生的主要分子基礎(chǔ),而氨基糖甙類抗生素的應(yīng)用增強(qiáng)了該突變的表型表達(dá);未能證實(shí)線粒體單體型以及核基因TRMU和MTO1對家系成員C1494T突變的表型具有修飾作用。
[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.
【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】：2011
【分類號】：R764
本文編號：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.
【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】：2011
【分類號】：R764
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 ;Mitochondrial rRNA and tRNA and hearing function[J];Cell Research;2007年03期
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