二叶式主动脉瓣畸形相关GATA5基因突变谱分析

摘要

Objective To analyze the mutational spectrum of GATA5 gene associated with congenital bicuspid aortic valve(BAV).Methods A total of 150 patients with congenital BAV and a total of 200 healthy subjects as controls were enrolled.The clinical data were collected,and the peripheral venous blood specimens were prepared.The genomic DNA was isolated by DNA purification kit.The coding regions and splice junction sites of GATA5 gene were amplified by polymerase chain reaction,and the amplicons were sequenced by di-deoxynucleotide chain termination technique.The acquired sequences were aligned with those of GATA5 released from GenBank to identify the likely sequence mutations.The online software MUSCLE and MutationTaster were used to evaluate whether the altered amino acids were conserved evolutionarily and whether the mutations were causative,respectively.Results Two novel heterozygous GATA5 mutations,p.M219I and p.T289I,were identified in 2 congenital BAV patients,respectively,with a mutational prevalence of approximately 1 .33%.The 2 mutations,which altered the amino acids completely conserved evolutionarily,were absent in the 200 controls and were both predicted to be pathogenic.Conclusions This study links novel GATA5 mutations to congenital BAV,which helps to reveal new molecular mechanism underlying congenital BAV.%目的：分析先天性二叶式主动脉瓣畸形（BAV）相关GATA5基因突变谱。方法收集150例先天性BAV患者和200名健康对照者的临床资料和血标本，使用DNA纯化试剂盒抽提基因组DNA。通过聚合酶链反应扩增GATA5基因的编码区和剪接点，采用双脱氧核苷链末端合成终止法测序，将所测序列与GenBank数据库中的GATA5基因序列进行比对以发现GATA5基因突变。分别应用在线软件MUSCLE和MutationTaster评估突变氨基酸的保守性和致病性。结果在2例先天性BAV患者各发现1个新的GATA5基因杂合突变，即p．M219I和p．T289I，突变率约为1．33％。这2种突变均不存在于200名健康对照者中。多序列比对显示被改变氨基酸在进化上均完全保守，致病性预测表明这2种突变均具有致病性。结论发现BAV相关GATA5基因新突变，有助于揭示BAV新的分子机制。