一先天性白内障家系的疾病相关候选基因定位

发布时间：2018-02-05 16:41

本文关键词： 先天性白内障基因突变剪切位点突变 CRYBA3/A1基因　出处：《浙江大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:先天性白内障是导致儿童视力损害甚至失明的主要眼病之一,其中常染色体显性遗传是遗传性先天性白内障的主要的遗传方式。本病的遗传学致病机制并非单一,迄今为止已发现超过40种不同的独立致病位点。本实验对一个中国先天性白内障家系进行了致病基因筛查研究。方法:本实验纳入了一个中国先天性白内障家系,纳入四代成员共12人,其中患者5人。所有家系成员均接受了相关病史调查和详细的眼科检查,包括视力检查、散瞳后裂隙灯及直接检眼镜检查等。采集每个家系成员的外周血并提取基因组DNA。应用聚合酶链式反应(PCR)扩增所有疾病相关候选基因的编码区及内含子、外显子交界区。对扩增产物进行双向测序,并分析基因序列。用限制性片段长度多态性分析(RFLP)验证突变,并应用多种生物信息学分析方法(Human Splicing Finder Matrices and MaxEnt tools,Annovar)预测突变对蛋白质功能的影响。结果:通过家系图绘制及分析,该家系的遗传方式被确定为常染色体显性遗传。直接测序发现家系中所有患者的CRYBA3/A1基因编码区一条等位基因的第2个外显子前第2位碱基发生了由A到G的改变(c.30-2AG),而该突变在家系中未患病成员和100名无亲缘关系的健康对照中均未发现。利用限制性片段长度多态性分析验证了该突变的存在,并利用数种在线生物信息学分析工具预测出该突变为可能致病突变。结论:本实验在一个中国先天性白内障的家系中发现了 CRYBA3/A1基因c.30-2 AG剪切位点突变,该突变被验证与先天性白内障存在共分离。进一步支持了CRYBA3/A1基因突变对于先天性白内障的致病性,并扩大了先天性白内障相关的致病突变谱。
[Abstract]:Objective: congenital cataract is one of the major eye diseases that lead to visual impairment or even blindness in children. Autosomal dominant inheritance is the main genetic mode of hereditary congenital cataract. The genetic mechanism of this disease is not single. To date, more than 40 different independent pathogenetic loci have been identified. A genetic screening study was conducted in a Chinese family of congenital cataract. A Chinese family of congenital cataract. 12 members were included in the four generations, including 5 patients. All the family members underwent relevant medical history investigation and detailed ophthalmic examination, including visual examination. Slit-lamp and direct ophthalmoscope examination after pupil dilatation. The peripheral blood of each family member was collected and genomic DNA was extracted. Polymerase chain reaction (PCR) was used. The coding regions and introns of all disease-related candidate genes were amplified. The amplification products were sequenced in two directions and the gene sequence was analyzed. The mutation was verified by restriction fragment length polymorphism (RFLP) analysis. A variety of bioinformatics analysis methods were used to analyze human Splicing Finder Matrices and MaxEnt tools. Annovar. was used to predict the effect of mutation on protein function. Results: family map was used to draw and analyze the mutation. The genetic pattern of the family was determined to be autosomal dominant inheritance. Direct sequencing revealed that the second base of the second exon of an allele in the CRYBA3/A1 gene coding region of all patients in the family was found to be at the second base. Changes from A to G (. C.30-2AGs). The mutation was not found in none of the family members and 100 unrelated healthy controls. The existence of the mutation was verified by restriction fragment length polymorphism (RFLP) analysis. Several online bioinformatics tools were used to predict the mutation as a possible pathogenic mutation. Conclusion: this study was found in a Chinese pedigrees with congenital cataract. CRYBA3/A1 gene c. 30-2 AG splicing site mutation. This mutation has been proved to be coisolated from congenital cataract, which further supports the pathogenicity of CRYBA3/A1 gene mutation in congenital cataract and expands the spectrum of pathogenicity mutation associated with congenital cataract.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R776.1

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