發(fā)布時(shí)間：2018-06-08 15:53

  本文選題：特發(fā)性全身性癲癇 + CACNA1H基因��； 參考：《廣西醫(yī)科大學(xué)》2012年博士論文

【摘要】：第一部分特發(fā)性全身性癲癇CACNA1H基因的突變分析 目的：探討T型鈣通道CACNA1H基因變異與中國(guó)廣西漢族兒童特發(fā)性全身性癲癇(IGE)的關(guān)系。 方法：對(duì)100例中國(guó)廣西漢族IGE患兒和100例漢族健康對(duì)照者應(yīng)用聚合酶鏈?zhǔn)椒磻?yīng)(Polymerase Chain Reaction,PCR)方法擴(kuò)增CACNA1H基因外顯子9～11編碼區(qū)及兩側(cè)各50個(gè)堿基以上的序列,PCR產(chǎn)物經(jīng)ABI-PRISM3730全自動(dòng)DNA測(cè)序儀自動(dòng)測(cè)序,然后與NCBI數(shù)據(jù)庫(kù)中CACNA1H基因的序列進(jìn)行比對(duì),了解CACNA1H基因突變情況,利用生物信息學(xué)方法預(yù)測(cè)CACNA1H基因突變對(duì)蛋白質(zhì)功能的影響。 結(jié)果： 1.在IGE患兒中,發(fā)現(xiàn)CACNA1H基因兩個(gè)新的雜合子突變,1254211GA及1254303AT,均位于第10外顯子上。分別導(dǎo)致所編碼的氨基酸發(fā)生改變,Arg735G1n及Arg766Trp。 2. CACNA1H基因編碼第735位及第766位Arg在生物進(jìn)化過(guò)程中處于部分保守區(qū)域。 3.用SIFT和PolyPhen程序預(yù)測(cè)CACNA1H基因點(diǎn)突變,顯示Arg766Trp有可能影響蛋白質(zhì)的功能。 4.共發(fā)現(xiàn)CACNA1H基因的14個(gè)多態(tài)性位點(diǎn),其中有5個(gè)變異在NCBI的SNP數(shù)據(jù)庫(kù)中未見(jiàn)收錄,在IGE組及對(duì)照組中比較了7個(gè)SNPs位點(diǎn)的基因型、等位基因及單體型的頻率分布,結(jié)果顯示無(wú)顯著性差異。 結(jié)論：T型鈣通道CACNA1H基因突變可能與中國(guó)廣西IGE患兒的發(fā)生有關(guān),CACNA1H基因很可能是IGE的易感基因。 第二部分GABRB3基因多態(tài)性與兒童特發(fā)性全身性癲癇相關(guān)性研究 目的：探討GABRB3基因多態(tài)性與中國(guó)廣西漢族兒童特發(fā)性全身性癲癇(IGE)的關(guān)系。 方法：以GABRB3基因上5個(gè)單核苷酸多態(tài)性(SNPs)rs4906902,rs981778,rs2059574,rs890317,rs1426217為遺傳標(biāo)記,運(yùn)用聚合酶鏈反應(yīng)-限制性片段長(zhǎng)度多態(tài)性及DNA測(cè)序法對(duì)100例廣西漢族IGE患兒及200例漢族健康對(duì)照者進(jìn)行5個(gè)SNPs位點(diǎn)的測(cè)定,應(yīng)用SPSS17.0軟件比較兩組兒童中各SNPs位點(diǎn)基因型頻率及等位基因頻率,用SHESIS軟件進(jìn)行單體型的構(gòu)建及分析。 結(jié)果：GABRB3基因中單個(gè)SNPs的基因型頻率及等位基因頻率在IGE組及對(duì)照組之間無(wú)顯著性差異。GABRB3基因單體型A-A-T-A-G(SNPs順序：rs4906902,rs981778,rs2059574,rs890317,rs1426217)在IGE組出現(xiàn)的頻率明顯高于正常對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義。 結(jié)論：γ一氨基丁酸A型受體亞單位GABRB3基因可能與中國(guó)廣西IGE患兒相關(guān),GABRB3基因可能是廣西IGE患兒的易感基因。
[Abstract]:Part I: mutation analysis of CACNA1H gene in idiopathic generalized epilepsy objective: to investigate the relationship between the variation of CACNA1H gene in T-type calcium channel and IGE in Han Chinese children of Guangxi Han nationality. Methods: 100 cases of Han nationality in Guangxi were studied. PCR products of CACNA1H gene exon 9M11 and 50 bases on both sides were amplified by polymerase chain reaction Polymerase chain reaction (Polymerase chain reaction) PCRmethod. The PCR products were sequenced by ABI-PRISM3730 automatic DNA sequencer. Then the sequence of CACNA1H gene was compared with the sequence of CACNA1H gene in NCBI database to understand the mutation of CACNA1H gene, and the effect of CACNA1H gene mutation on protein function was predicted by bioinformatics. Results: 1. Two new heterozygous mutations of CACNA1H gene, 1254211GA and 1254303AT, were found in children with IGE, both at exon 10. The results showed that the encoded amino acids changed Arg735G1n and Arg766TRP. 2, respectively. CACNA1H gene encodes 735th and 766-bit Arg in a partially conserved region during biological evolution. The point mutation of CACNA1H gene was predicted by sift and PolyPhen program, which indicated that Arg766Trp might affect the function of protein. 4. A total of 14 polymorphic loci of CACNA1H gene were found, 5 of which were not included in the SNP database of NCBI. Genotypes, alleles and haplotypes of 7 SNPs loci were compared in the IGE group and the control group. Conclusion the mutation of CACNA1H gene may be related to the occurrence of IGE in Guangxi, China. The second part of GABRB3 gene polymorphism is associated with idiopathic systemic IGE in children. Objective: to investigate the relationship between GABRB3 gene polymorphism and IGE in Chinese Han nationality children with idiopathic generalized epilepsy. Methods: five single nucleotide polymorphisms (SNPs4906902rs981778 rs2059574rs890317rs1426217) of GABRB3 gene were used as genetic markers. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing were used to detect 5 SNPs loci in 100 children with IGE in Guangxi Han nationality and 200 healthy controls. SPSS 17.0 software was used to compare the genotype frequency and allele frequency of each SNPs locus between the two groups. Results there was no significant difference in genotype frequency and allele frequency of single SNPs between IGE group and control group. The order of haplotype A-T-A-SNPs of GABRB3 gene was: rs4906902rs981778rs2059574rs890317rs1426217). The rate was significantly higher than that in the normal control group. Conclusion: the GABRB3 gene of 緯 -aminobutyric acid type A receptor subunit may be the susceptible gene of IGE children in Guangxi, China.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R742.1

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本文編號(hào)：1996338