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目的对1例遗传性多发性骨软骨瘤患者的EXT1和EXT2基因编码序列进行突变检测,寻找致病突变。方法采集患者及其家系成员外周血,应用PCR技术扩增EXT1和EXT2基因编码区及外显子-内含子交界区,再对产物进行测序。结果发现EXT1基因有7种内含子突变(c.1165-156G>GT、c.1284+66G>GA、c.1536+59G>GT、c.1536+91_99del AGGCTCCCC、c.1537-51A>AG、c.1722+202T>TC、c.1723-103C>CG),发现EXT2基因有6种内含子突变(c.70-80A>AC、c.636-79A>AC、c.1179-18T>TA、c.1595-195C>T、c.1905-51T>TC、c.2035-41T>TC)、1种外显子突变(c.138G>GA,p.E28K)。其中c.1536+91_99del AGGCTCCCC和p.E28K两种突变是新发现的多态位点,其它12种突变均为人类基因突变数据库收率的多态位点。结论未能找到该家系的明确致病基因,该家系的发病是否由EXT1和EXT2基因以外的EXT基因家族突变引起还需进一步的连锁定位分析。
Objective To detect the mutation of EXT1 and EXT2 gene coding sequence in one case of hereditary multiple osteochondroma to find the pathogenic mutation. Methods Peripheral blood samples of patients and their pedigree were collected. The coding region of EXT1 and EXT2 gene and exon-intron junction were amplified by PCR and the products were sequenced. As a result, there were 7 intronic mutations in the EXT1 gene (c.1165-156G> GT, c.1284 + 66G> GA, c.1536 + 59G> GT, c.1536 + 91_99del AGGCTCCCC, c.1537-51A> AG , C.1722 + 202T> TC, c.1723-103C> CG). There were 6 intronic mutations in the EXT2 gene (c.70-80A> AC, c.636-79A> AC, c.1179-18T > TA, c.1595-195C> T, c.1905-51T> TC, c.2035-41T> TC), one exon mutation (c.138G> GA, p.E28K). Which c.1536 +91_99del AGGCTCCCC and p.E28K two mutations are newly discovered polymorphic sites, the other 12 kinds of mutations are human genetic mutation database yield polymorphism. Conclusion No specific pathogenic genes were found in this pedigree. Whether the incidence of this pedigree was caused by mutations in the EXT gene family other than EXT1 and EXT2 genes requires further linkage analysis.