本文關(guān)鍵詞： 序列特異性引物聚合酶鏈?zhǔn)椒磻?yīng)(PCR-SSP) 先天性白內(nèi)障 CRYBB1 基因突變　出處：《新鄉(xiāng)醫(yī)學(xué)院》2012年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：背景基于本課題組于前期實(shí)驗(yàn)研究中,對(duì)一常染色體顯性遺傳性先天性白內(nèi)障家系候選基因進(jìn)行基因組全序列測(cè)序時(shí)發(fā)現(xiàn)CRYBB1晶體蛋白基因第6外顯子的剪切位點(diǎn)(AG)處發(fā)生G→A基因位點(diǎn)突變,為進(jìn)一步研究此突變的性質(zhì)和意義,本實(shí)驗(yàn)選用臨床表現(xiàn)型相似的2個(gè)常染色體顯性遺傳性先天性白內(nèi)障家系,篩檢該2個(gè)家系與此突變位點(diǎn)的關(guān)系。 目的采用序列特異性引物聚合酶鏈?zhǔn)椒磻?yīng)(PCR-SSP)法,對(duì)2個(gè)常染色體顯性遺傳性先天性白內(nèi)障家系進(jìn)行研究,篩檢其是否存在CRYBBl晶體蛋白基因第6外顯子剪切位點(diǎn)(AG)處發(fā)生的G→A基因突變,進(jìn)一步分析判定此G→A基因突變的性質(zhì)。 方法收集研究對(duì)象,提取基因組DNA,采用PCR-SSP法,根據(jù)CRYBB1晶體蛋白基因第6外顯子剪切位點(diǎn)(AG)處發(fā)生G→A基因突變,針對(duì)此基因突變?cè)O(shè)計(jì)相應(yīng)的序列特異性引物引導(dǎo)聚合酶鏈?zhǔn)椒磻?yīng),然后根據(jù)PCR擴(kuò)增產(chǎn)物的電泳結(jié)果,判斷其各自的DNA基因型,驗(yàn)證分析該2個(gè)常染色體顯性遺傳性先天性白內(nèi)障家系是否發(fā)生此G→A基因突變。 結(jié)果本實(shí)驗(yàn)研究中的所有的研究對(duì)象CRYBBl晶體蛋白基因第6外顯子剪切位點(diǎn)(AG)處均未出現(xiàn)G→A基因突變,其基因型為純合子GG型。 結(jié)論CRYBB1晶體蛋白基因第6外顯子剪切位點(diǎn)(AG)處發(fā)生的G→A基因突變?yōu)榕及l(fā)的點(diǎn)突變,此基因突變不具有基因突變的多態(tài)性和廣泛性。PCR-SSP法是一種可靠的有效的在人群中篩檢基因點(diǎn)突變的可行性方法。
[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. 鈫捍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.
【學(xué)位授予單位】：新鄉(xiāng)醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R776.1

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