動(dòng)脈導(dǎo)管未閉兒童TFAP2B基因突變篩查
發(fā)布時(shí)間：2019-04-11 20:17
【摘要】：目的 動(dòng)脈導(dǎo)管未閉(PDA)是小兒常見(jiàn)的先天性心臟病,遺傳因素在PDA的發(fā)生中占重要地位,尋找PDA發(fā)病相關(guān)基因已成為當(dāng)前心血管領(lǐng)域研究的熱點(diǎn)。心臟特異轉(zhuǎn)錄因子TFAP2B在神經(jīng)嵴細(xì)胞分化及動(dòng)脈導(dǎo)管的形成過(guò)程中發(fā)揮重要作用,本研究旨在探討PDA兒童是否存在TFAP2B基因突變。 研究對(duì)象與方法 選取74例散發(fā)的單純性PDA兒童作為病例組,以100例健康兒童作為對(duì)照組,分別取外周血提取基因組DNA。設(shè)計(jì)TFAP2B基因的7對(duì)外顯子及其兩側(cè)部分內(nèi)含子的引物,應(yīng)用聚合酶鏈反應(yīng)(PCR)擴(kuò)增目的基因。PCR擴(kuò)增完畢后,將產(chǎn)物放入2%瓊脂糖凝膠中進(jìn)行電泳,檢測(cè)合格后進(jìn)行純化。純化產(chǎn)物采用雙脫氧核苷鏈末端合成終止法自動(dòng)測(cè)序,將所測(cè)得序列與NCBI及Genbank數(shù)據(jù)庫(kù)中TFAP2B基因序列進(jìn)行比對(duì),分析TFAP2B基因有無(wú)突變或多態(tài)性。 結(jié)果 1.PDA兒童TFAP2B基因測(cè)序結(jié)果中,未發(fā)現(xiàn)編碼區(qū)域的錯(cuò)義突變。 2.病例組中26例兒童TFAP2B基因存在1個(gè)新的單核苷酸多態(tài)性位點(diǎn),編碼區(qū)上游第34位鳥(niǎo)嘌呤(G)轉(zhuǎn)換為腺嘌呤(A),即c.1-34G→A多態(tài)。對(duì)照組中32例兒童存在相同的多態(tài)性位點(diǎn),該位點(diǎn)在NCBI及Genbank數(shù)據(jù)庫(kù)中未見(jiàn)報(bào)道。通過(guò)多態(tài)性分析,該位點(diǎn)的3種基因型GG、AG、AA在總體研究人群中的頻率分布符合Hardy-Weinberg遺傳平衡(x=0.129,P=0.938)。等位基因G、A及基因型GG、AG、AA在病例組和對(duì)照組間的頻率分布無(wú)差異(P均0.05) 結(jié)論 1.本研究未發(fā)現(xiàn)動(dòng)脈導(dǎo)管未閉兒童存在TFAP2B基因突變。 2.74例動(dòng)脈導(dǎo)管未閉兒童中26例和100例健康兒童中32例在TFAP2B基因編碼區(qū)上游第34位堿基均存在一個(gè)新的單核苷酸多態(tài)性位點(diǎn),鳥(niǎo)嘌呤(G)轉(zhuǎn)換為腺嘌呤(A),即c.1-34G→A多態(tài),提示TFAP2B基因c.1-34G→A多態(tài)位點(diǎn)與動(dòng)脈導(dǎo)管未閉的發(fā)生可能無(wú)關(guān)。
[Abstract]:Aim: patent ductus arteriosus (PDA) is a common congenital heart disease in children. Genetic factors play an important role in the occurrence of PDA. It has become a hot topic in the field of cardiovascular research to search for genes related to the pathogenesis of PDA. Heart-specific transcription factor TFAP2B plays an important role in the differentiation of neural crest cells and the formation of ductus arteriosus. The aim of this study was to investigate the existence of TFAP2B gene mutation in children with PDA. Subjects and methods 74 children with sporadic simple PDA were selected as the case group and 100 healthy children as the control group. The genomic DNA. was extracted from the peripheral blood. The primers of exon 7 and its introns of TFAP2B gene were designed and amplified by polymerase chain reaction (PCR). After PCR amplification, the product was placed into 2% agarose gel for electrophoresis and purified. The purified products were sequenced automatically by the method of terminal synthesis of dideoxynucleoside chain. The sequences were compared with those of TFAP2B gene in NCBI and Genbank database, and the mutation or polymorphism of TFAP2B gene was analyzed. Results No missense mutation in the coding region was found in TFAP2B gene sequencing of 1.PDA children. 2. In the case group, there was a new single nucleotide polymorphism site in the TFAP2B gene of 26 children. Guanine (G) was transformed into adenine (A), at the 34th position upstream of the coding region, that is, c. 1 / 34 G / A polymorphism. The same polymorphism was found in 32 children in the control group, which was not reported in NCBI and Genbank databases. The polymorphism analysis showed that the frequency distribution of the three genotypes of GG,AG,AA in the population was consistent with the genetic balance of Hardy-Weinberg (x = 0.129, P = 0.938). There was no difference in frequency distribution of allele G, A and genotype GG,AG,AA between case group and control group (P 0.05) conclusion 1. No mutations in TFAP2B gene were found in children with patent ductus arteriosus. In 2.74 cases of patent ductus arteriosus, 26 cases of patent ductus arteriosus and 32 cases of healthy children had a new single nucleotide polymorphism site at 34bp upstream of the coding region of TFAP2B gene. Guanine (G) was converted to adenine (A),. That is, c. 1-34-G / A polymorphism suggests that the polymorphism of c. 1-34-G / A of TFAP2B gene may not be related to the occurrence of patent ductus arteriosus (ductus arteriosus).
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R725.4
本文編號(hào)：2456715
[Abstract]:Aim: patent ductus arteriosus (PDA) is a common congenital heart disease in children. Genetic factors play an important role in the occurrence of PDA. It has become a hot topic in the field of cardiovascular research to search for genes related to the pathogenesis of PDA. Heart-specific transcription factor TFAP2B plays an important role in the differentiation of neural crest cells and the formation of ductus arteriosus. The aim of this study was to investigate the existence of TFAP2B gene mutation in children with PDA. Subjects and methods 74 children with sporadic simple PDA were selected as the case group and 100 healthy children as the control group. The genomic DNA. was extracted from the peripheral blood. The primers of exon 7 and its introns of TFAP2B gene were designed and amplified by polymerase chain reaction (PCR). After PCR amplification, the product was placed into 2% agarose gel for electrophoresis and purified. The purified products were sequenced automatically by the method of terminal synthesis of dideoxynucleoside chain. The sequences were compared with those of TFAP2B gene in NCBI and Genbank database, and the mutation or polymorphism of TFAP2B gene was analyzed. Results No missense mutation in the coding region was found in TFAP2B gene sequencing of 1.PDA children. 2. In the case group, there was a new single nucleotide polymorphism site in the TFAP2B gene of 26 children. Guanine (G) was transformed into adenine (A), at the 34th position upstream of the coding region, that is, c. 1 / 34 G / A polymorphism. The same polymorphism was found in 32 children in the control group, which was not reported in NCBI and Genbank databases. The polymorphism analysis showed that the frequency distribution of the three genotypes of GG,AG,AA in the population was consistent with the genetic balance of Hardy-Weinberg (x = 0.129, P = 0.938). There was no difference in frequency distribution of allele G, A and genotype GG,AG,AA between case group and control group (P 0.05) conclusion 1. No mutations in TFAP2B gene were found in children with patent ductus arteriosus. In 2.74 cases of patent ductus arteriosus, 26 cases of patent ductus arteriosus and 32 cases of healthy children had a new single nucleotide polymorphism site at 34bp upstream of the coding region of TFAP2B gene. Guanine (G) was converted to adenine (A),. That is, c. 1-34-G / A polymorphism suggests that the polymorphism of c. 1-34-G / A of TFAP2B gene may not be related to the occurrence of patent ductus arteriosus (ductus arteriosus).
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R725.4
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 呂瑛;張密林;解啟蓮;高磊;;動(dòng)脈導(dǎo)管未閉發(fā)病機(jī)制研究進(jìn)展[J];實(shí)用兒科臨床雜志;2007年13期
，本文編號(hào)：2456715
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