PCR-SSP法筛检2个先天性白内障家系CRYBB1第6外显子基因位点突变

发布时间：2018-01-21 17:39

本文关键词： 序列特异性引物聚合酶链式反应(PCR-SSP) 先天性白内障 CRYBB1 基因突变　出处：《新乡医学院》2012年硕士论文　论文类型：学位论文

【摘要】：背景基于本课题组于前期实验研究中,对一常染色体显性遗传性先天性白内障家系候选基因进行基因组全序列测序时发现CRYBB1晶体蛋白基因第6外显子的剪切位点(AG)处发生G→A基因位点突变,为进一步研究此突变的性质和意义,本实验选用临床表现型相似的2个常染色体显性遗传性先天性白内障家系,筛检该2个家系与此突变位点的关系。目的采用序列特异性引物聚合酶链式反应(PCR-SSP)法,对2个常染色体显性遗传性先天性白内障家系进行研究,筛检其是否存在CRYBBl晶体蛋白基因第6外显子剪切位点(AG)处发生的G→A基因突变,进一步分析判定此G→A基因突变的性质。方法收集研究对象,提取基因组DNA,采用PCR-SSP法,根据CRYBB1晶体蛋白基因第6外显子剪切位点(AG)处发生G→A基因突变,针对此基因突变设计相应的序列特异性引物引导聚合酶链式反应,然后根据PCR扩增产物的电泳结果,判断其各自的DNA基因型,验证分析该2个常染色体显性遗传性先天性白内障家系是否发生此G→A基因突变。结果本实验研究中的所有的研究对象CRYBBl晶体蛋白基因第6外显子剪切位点(AG)处均未出现G→A基因突变,其基因型为纯合子GG型。结论CRYBB1晶体蛋白基因第6外显子剪切位点(AG)处发生的G→A基因突变为偶发的点突变,此基因突变不具有基因突变的多态性和广泛性。PCR-SSP法是一种可靠的有效的在人群中筛检基因点突变的可行性方法。
[Abstract]:Background based on our research group in the previous experimental study. Sequencing of candidate genes from an autosomal dominant congenital cataract pedigree revealed that G occurred at the cleavage site of exon 6 of CRYBB1 crystal protein gene. 鈫扞n order to further study the character and significance of A gene mutation, two families with autosomal dominant congenital cataract with similar clinical manifestations were selected in this study. The relationship between the two families and the mutation site was screened. Objective to study two pedigrees with autosomal dominant congenital cataract by sequence specific primer polymerase chain reaction (PCR-SSP). Screening for the presence of G at the cleavage site of exon 6 of the CRYBBl crystal protein gene 鈫扢utation of A gene, further analysis and determination of the G. 鈫扵he nature of A gene mutation. Methods Genomic DNA was extracted from CRYBB1 crystal protein gene, and G was generated according to the cleavage site of exon 6 of CRYBB1 crystal protein gene by PCR-SSP method. 鈫扐ccording to the mutation of A gene, the sequence specific primers were designed to guide the polymerase chain reaction, and then according to the electrophoresis results of PCR amplification products, their respective DNA genotypes were determined. To verify the occurrence of this G in the two autosomal dominant congenital cataract families. 鈫扐 gene mutation. Results none of the subjects in this study had G at exon 6 of CRYBBl crystal protein gene. 鈫扵he genotype of A gene was homozygous GG type. Conclusion G at the cleavage site of exon 6 of CRYBB1 crystal protein gene 鈫扵he mutation of A gene is an accidental point mutation. This mutation does not have the polymorphism of gene mutation and the extensibility. PCR-SSP method is a reliable and effective method for screening point mutation of gene in the population.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R776.1

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