CAJ | 학술논문

目的筛查先天性房间隔缺损相关GATA4基因的新突变.方法收集85例先天性房间隔缺损患者和200名健康者的临床资料及外周静脉血标本.应用聚合酶链反应扩增GATA4基因的全部外显子,采用双脱氧核苷链末端合成终止法对全部扩增片段进行测序.以BLAST程序将所测序列与GenBank中的已知序列进行比对,以识别基因突变.采用Clustal W软件分析突变氨基酸的保守性.结果在其中3例先天性房间隔缺损患者的GATA4基因各识别出1个新的杂合错义突变,即第267、354和407位的密码子分别由GTG、ACC和CCA变为ATG、GCC和CAA,亦即c.799G>A、c.1060A>G和c.1220C>A突变,相应地导致第267、354和407位的氨基酸分别由缬氨酸、苏氨酸和脯氨酸变为蛋氨酸、丙氨酸和谷氨酰胺,也称为p.V267M、p.T354A和p.P407Q突变.200名健康对照者均无上述突变.多物种GATA4序列比对显示,第267位的缬氨酸和第407位的脯氨酸在进化上完全保守.结论在先天性房间隔缺损患者的GATA4基因识别出3个新的杂合错义突变,揭示了先天性房间隔缺损新的分子病因,有助于先天性房间隔缺损的早期防治.
목적 사사선천성방간격결손상관GATA4기인적신돌변.방법 수집85례선천성방간격결손환자화200명건강자적림상자료급외주정맥혈표본.응용취합매련반응확증GATA4기인적전부외현자,채용쌍탈양핵감련말단합성종지법대전부확증편단진행측서.이BLAST정서장소측서렬여GenBank중적이지서렬진행비대,이식별기인돌변.채용Clustal W연건분석돌변안기산적보수성.결과 재기중3례선천성방간격결손환자적GATA4기인각식별출1개신적잡합착의돌변,즉제267、354화407위적밀마자분별유GTG、ACC화CCA변위ATG、GCC화CAA,역즉c.799G>A、c.1060A>G화c.1220C>A돌변,상응지도치제267、354화407위적안기산분별유힐안산、소안산화포안산변위단안산、병안산화곡안선알,야칭위p.V267M、p.T354A화p.P407Q돌변.200명건강대조자균무상술돌변.다물충GATA4서렬비대현시,제267위적힐안산화제407위적포안산재진화상완전보수.결론 재선천성방간격결손환자적GATA4기인식별출3개신적잡합착의돌변,게시료선천성방간격결손신적분자병인,유조우선천성방간격결손적조기방치.
Objective To screen the gene CATA4 for novel mutations associated with congenital atria] septal defect ( ASD). Methods The clinical data and peripheral venous blood specimen from 85 unrelated subjects with congenital ASD were collected and analyzed in contrast to 200 healthy individuals. The coding exons and the exon/intron boundaries of GATA4 gene were amplified by polymerase chain reaction and sequenced using the di-deoxynucleotide chain termination procedure. The obtained sequences were aligned with those publicized in GenBank with the help of programme BLAST to identify the sequence variations. The software Clustal W was applied to analysis of the conservation of altered amino acids. Results Three novel heterozygous missense GATA4 mutations were identified in 3 of 85 ASD patients, respectively. Namely, the triplet substitutions of ATG for GTG at codon 267, GCC for ACC at codon 354, and CAA for CCA at codon 407, predicting the conversions of valine into methionine at amino acid residue 267 ( V267M) , threonine into alanine at amino acid residue 354 (T354A), and proline into glutamine at amino acid residue 407 (P407Q) , were identified. No mutation was detected in 200 healthy controls. A cross-species alignment of GATA4 encoded protein sequences showed that the valine at amino acid residue 267 and proline at amino acid residue 407 were completely conserved evolutionarily. Conclusion Three novel heterozygous missense GATA4 mutations were identified in patients with congenital ASD, which reveals new molecular etiology responsible for ASD, and contributes to the early prophylaxis and therapy for ASD.