动脉导管未闭儿童TFAP2B基因突变筛查
发布时间：2019-04-11 20:17
【摘要】：目的 动脉导管未闭(PDA)是小儿常见的先天性心脏病,遗传因素在PDA的发生中占重要地位,寻找PDA发病相关基因已成为当前心血管领域研究的热点。心脏特异转录因子TFAP2B在神经嵴细胞分化及动脉导管的形成过程中发挥重要作用,本研究旨在探讨PDA儿童是否存在TFAP2B基因突变。 研究对象与方法 选取74例散发的单纯性PDA儿童作为病例组,以100例健康儿童作为对照组,分别取外周血提取基因组DNA。设计TFAP2B基因的7对外显子及其两侧部分内含子的引物,应用聚合酶链反应(PCR)扩增目的基因。PCR扩增完毕后,将产物放入2%琼脂糖凝胶中进行电泳,检测合格后进行纯化。纯化产物采用双脱氧核苷链末端合成终止法自动测序,将所测得序列与NCBI及Genbank数据库中TFAP2B基因序列进行比对,分析TFAP2B基因有无突变或多态性。 结果 1.PDA儿童TFAP2B基因测序结果中,未发现编码区域的错义突变。 2.病例组中26例儿童TFAP2B基因存在1个新的单核苷酸多态性位点,编码区上游第34位鸟嘌呤(G)转换为腺嘌呤(A),即c.1-34G→A多态。对照组中32例儿童存在相同的多态性位点,该位点在NCBI及Genbank数据库中未见报道。通过多态性分析,该位点的3种基因型GG、AG、AA在总体研究人群中的频率分布符合Hardy-Weinberg遗传平衡(x=0.129,P=0.938)。等位基因G、A及基因型GG、AG、AA在病例组和对照组间的频率分布无差异(P均0.05) 结论 1.本研究未发现动脉导管未闭儿童存在TFAP2B基因突变。 2.74例动脉导管未闭儿童中26例和100例健康儿童中32例在TFAP2B基因编码区上游第34位碱基均存在一个新的单核苷酸多态性位点,鸟嘌呤(G)转换为腺嘌呤(A),即c.1-34G→A多态,提示TFAP2B基因c.1-34G→A多态位点与动脉导管未闭的发生可能无关。
[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).
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.4
本文编号：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).
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.4
【参考文献】
相关期刊论文 前1条
1 吕瑛;张密林;解启莲;高磊;;动脉导管未闭发病机制研究进展[J];实用儿科临床杂志;2007年13期
，本文编号：2456715
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