低钾型周期性麻痹家族基因序列分析

发布时间：2018-07-11 11:33

本文选题：家族性低钾型周期性麻痹 + 离子通道　；参考：《吉林大学》2014年硕士论文

【摘要】：实验目的：随着科技的发展，人们对疾病的认识逐渐深入，达到分子水平。离子通道病的概念走入人们的视线并逐渐被了解。家族性低钾型周期性麻痹是最常见的离子通道病之一，它是一种常染色体显性遗传病，临床上以骨骼肌反复发作弛缓性无力及发作时血清钾降低为主要特征，通常由精神紧张、寒冷、感染、输入葡萄糖溶液、体温下降、代谢性碱中毒、麻醉以及应用类固醇药物等诱发，给予补钾治疗后症状可基本消失。目前临床上主要依据发病形式及发病时患者血清钾浓度对该病进行诊断与治疗。本实验通过对一个家族性周期性麻痹家系成员进行基因序列分析探讨其突变位点，研究其对临床的指导意义。材料与方法：实验对象为目标家系同一代中的6名成员，其中3人患有低钾型周期性麻痹，均为男性，发病形式基本相同，取其血样提取出全基因组DNA。查阅相关文献可知，家族性低钾型周期性麻痹患者大约55%到70%左右是由骨骼肌电压依赖性钙通道CACNA1S基因突变所致，约8%到10%是由电压门控性钠通道SCN4A基因突变所致。在国内，主要有CACNA1S的R528G、R1239H、H916Q以及SCN4A的R672H/C基因突变型，分别存在于CACNA1S基因外显子11、外显子30及SCN4A外显子12上。本实验主要针对上述三段序列进行序列分析。根据CACNA1S外显子11、30和SCN4A外显子12序列的上下游内含子序列设计引物，利用PCR技术，获得目的片段并进行体外扩增，最后对所获得产物进行测序。通过家族中HOKPP患者基因序列与正常家族成员基因序列及正常人基因序列相比较，明确HOKPP患者是否存在基因突变以及基因突变类型。结果：本实验最终结果为未在此家族中成员中发现有CACNA1S基因及SCN4A基因突变。分析可能的原因如下：①家族成员中HOKPP患者均为散发病例；②在本实验中，，仅对CACNAS1基因的30号外显子、11号外显子及SCN4A基因的12号外显子进行测序，该家族成员中HOKPP患者可能存在其他基因型突变；③并不是所有的家族性HOKPP患者都存在有基因突变，目前约有20%家族性低钾型周期性麻痹患者未能检测到突变基因。
[Abstract]:Objective: with the development of science and technology, people's understanding of disease has gradually deepened and reached the molecular level. The concept of ion channel disease came into the eye and was gradually understood. Familial hypokalemic periodic paralysis is one of the most common ion channel diseases. It is an autosomal dominant disorder characterized by recurrent flaccid weakness in skeletal muscle and reduction of serum potassium during attack. Cold, infection, infusion of glucose solution, hypothermia, metabolic alkalosis, anaesthesia and steroid use, etc. At present, the diagnosis and treatment of the disease are mainly based on the form of the disease and the serum potassium concentration of the patients. In this study, the mutation site of a family member with familial periodic paralysis was analyzed by gene sequence analysis, and its clinical significance was studied. Materials and methods: the subjects were 6 members of the same generation of target families. Among them, 3 of them were suffering from hypokalemic periodic paralysis, all of them were male, the form of the disease was basically the same, and the whole genome DNA was extracted from their blood samples. About 55% to 70% of familial hypokalemic periodic paralysis patients were caused by CACNA1S gene mutation in skeletal muscle voltage-dependent calcium channel, and about 8% to 10% were caused by SCN4A gene mutation in voltage-gated sodium channel. In China, CACNA1S R528GN R1239H / H916Q and SCN4A R672H / C gene mutation were found in CACNA1S exon 11, exon 30 and SCN4A exon 12, respectively. The aim of this experiment is to analyze the sequence of the above three segments. Primers were designed according to the upstream and downstream intron sequences of CACNA1S exon 1130 and SCN4A exon 12. The target fragments were obtained by PCR and amplified in vitro. Finally, the obtained products were sequenced. By comparing the gene sequences of HOKPP patients with those of normal family members and normal individuals, we can find out whether or not there is gene mutation and the type of gene mutation in patients with HOKPP. Results: no mutation of CACNA1S gene and SCN4A gene was found in this family. In this study, exon 30, exon 11 and exon 12 of CACNAS1 gene and SCN4A gene were only sequenced. Not all patients with familial HOKPP had gene mutations, and about 20% of patients with familial hypokalemic periodic paralysis could not detect mutation genes.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R746.3

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