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建立蛋白截短检测技术,分析家族性腺瘤样息肉病(FAP)发病相关基因APC基因的胚系突变,研究基因突变型与FAP疾病表型的关系。经临床诊断的22例FAP患者及43例散发性大肠癌患者,分别取外周静脉血和正常结肠粘膜组织,常规提取基因组DNA。应用蛋白截短检测技术,分段分析APC基因巨大的第15外显子,对检出截短蛋白的样本进行测序,以确定突变位点及突变性质。22例FAP患者中,5例患者存在APC基因第15外显子的胚系突变,均为碱基缺失造成的移码突变,导致截短蛋白的产生;43例散发性大肠癌患者中未检测到APC基因第15外显子的胚系突变。蛋白截短检测技术是一种高效的基因突变分析技术,适用于大片段基因(如APC基因第15外显子)截短型突变的检测,可作为FAP症状出现前的常规基因诊断技术的一项有效补充。
To establish a protein truncated detection technique to analyze the germline mutations of the APC gene associated with the pathogenesis of familial adenomatous polyposis (FAP) and to study the relationship between the gene mutation and the phenotype of FAP. Twenty-two patients with FAP and 43 patients with sporadic colorectal cancer were diagnosed clinically. Peripheral venous blood and normal colonic mucosa tissue were collected and genomic DNA was extracted routinely. Using the truncated protein detection technology, the exon 15 of APC gene was segmented and the truncated protein samples were sequenced to determine the mutation sites and the nature of mutations. Among the 22 patients with FAP, 5 patients had germline mutation of exon 15 of APC gene, all of which were frameshift mutations caused by base deletion, resulting in the production of truncated protein; 43 patients with sporadic colorectal cancer were not detected Germline mutation to exon 15 of APC gene. Protein truncation detection is a highly efficient technique for gene mutation analysis, which is suitable for the detection of truncated mutations in large fragment genes (such as exon 15 of APC gene), which can be used as one of the conventional gene diagnostic techniques before the appearance of FAP symptoms Effective supplement.