单纯腭裂并全面性癫伴热性惊厥附加症家系的基因定位及GABRD基因测序研究  

Gene Mapping and Candidate Gene GABRD Sequencing Analysis in Cleft Palate and Generalized Epilepsy with Febrile Seizures Plus Family

作　　者：谢晓华[1] 李丽芳 黄希顺[3] 尹景岗[4] Michael Y.Yin Robyn H.Wallace 郭学鹏[5] 王家勤[5] 

机构地区：[1]河南省人民医院口腔科,郑州450000 [2]加拿大渥太华卫生研究院 [3]郑州大学第一附属医院神经科,郑州450052 [4]美国田纳西大学医学科学中心,tn38163 [5]新乡医学院第三附属医院儿科,河南新乡453003

出　　处：《实用儿科临床杂志》2010年第20期1576-1579,共4页Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金(30370502)

摘　　要：目的对单纯腭裂并全面性癫伴热性惊厥附加症家系进行基因定位,并对候选基因GABRD进行测序研究。方法对家系进行全基因组扫描两点间连锁分析,设计GABRD基因9对外显子-内含子交界处内含子引物,PCR扩增患者及健康对照GABRD基因组序列,测序采用Sanger双脱氧链终止法,PCR产物直接测序,对比分析患者的突变序列及健康对照序列。结果两点连锁分析在1p36区域最大LOD值为1.68。GABRD基因测序显示第4外显子患者的序列分别为T/C和T/T,健康对照的序列分别为T/T和A/C。编码序列分析GGT、GGC及GGA均为甘氨酸。患者GABRD基因mRNA碱基序列为C425T多态性。GABRD第7外显子患者的序列分别为C/T和T/T,健康对照分别为T/T和C/C;异常序列位于mRNA第911碱基处,编码分析AGT及AGC均编码丝氨酸。患者的碱基序列出现了T911C多态性。结论单纯腭裂并全面性癫伴热性惊厥附加症致病基因在1p36区域取得一定的连锁关系,在该家系未发现突变基因;所发现的C425T和T911C单个碱基多态性为其他腭裂及癫家系的连锁分析、关联分析及分子遗传学研究提供了依据。Objective Gene mapping of cleft palate only and generalized epilepsy with febrile seizures plus pedigree,and sequencing analysis of the candidate gene GABRD.Methods Genome-wide linkage screen by two-point linkage analysis was used in this pedigree.Primers of intron-exon border were designed for all 9 exons of GABRD,and PCR products from patients and controls were directly sequenced.Mutant sequences from patients were compared to the controls.Results The highest two-point linkage analysis LOD score was 1.68 at 1p36.In GABRD mRNA 425 bp or exon 4,the sequences of patients were T/C and T/T,in normal controls were T/T and A/C,respectively.GGT,GGC and GGA were all coding amino acid Gly.The sequence difference was a C425T single nucleotide polymorphism（SNP）.In mRNA 911 bp or exon 7,the sequences in patients were C/T and T/T,but T/T and C/C in controls.Both AGT and AGC were coding amino acid Ser,it was a T911C SNP.Conclusions A positive linkage result was achieved from this pedigree at 1p36,no obvious mutant sequence was detected from the patients.The GABRD gene C425T and T911C SNP were useful for other gene mapping and gene association analysis in cleft palate and epilepsy pedigrees.

关 键 词：腭裂 癫 连锁分析 基因 单个碱基多态性 

分 类 号：R782.22[医药卫生—口腔医学] R742.1[医药卫生—临床医学]