多重PCR在瘢痕疙瘩Fas基因突变分析中的应用  

Multiplex PCR in analysis of Fas gene derived from human keloids

作　　者：靳睿[1] 高建华[1] 刘晓军[1] 

机构地区：[1]解放军第一军医大学南方医院整形外科,广东省广州市510515

出　　处：《中国临床康复》2003年第23期3190-3191,T004,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金(39870807)

摘　　要：目的:建立瘢痕疙瘩Fas基因常见突变外显子的多重PCR扩增反应体系,以利进行批量瘢痕疙瘩基因突变的筛选和检测。方法:取正常皮肤、增生瘢痕和瘢痕疙瘩组织DNA样本,根据瘢痕疙瘩Fas基因易发突变的外显子6,8,9之序列,按照多重PCR引物设计原则,建立3对相应引物,先分别找出各外显子的最佳扩增条件,再进行综合分析,最终找到一个理想的多重PCR扩增条件。结果:各外显子单一扩增片段和多重PCR扩增条带均清晰,产量较高,无非特异性扩增,产物长度与理论值一致;DNA测序证实,各扩增产物即各外显子基因片段。结论:成功建立了瘢痕疙瘩Fas基因常发突变外显子的多重PCR扩增体系,与以往单基因分次扩增相比,实现了一次同时扩增3个基因片段,为瘢痕疙瘩基因诊断和进一步相关的分子生物学研究提供了一个经济、快捷的方法,对于瘢痕疙瘩Fas基因的突变研究具有较高的应用价值。AIM:To establish a multiple polymerase chain reaction technique for detection and screening of Fas gene mutations in keloids in batches.METHODS: The whole sequences of exon 6,8 and the gene fragment of death domain of exon 9 of Fas gene from normal skins, hypertrophic scars and keloids of human were chosen and three pairs of primers were designed. After each fragment was amplified successfully, the appropriate condition of multiplex PCR was determined. RESULTS: The single amplified fragment of exon and multiplex PCR amplified bands of three products were seen in agarose gel electrophoresis clearly and uniquely, without nonspecific amplified fragment. The DNA sequences of these three bands indicated that the lengths coincided with the sequences, which had been predicted.

关 键 词：PCR 瘢痕疙瘩 Fas基因突变 应用 分子生物学 

分 类 号：R619.6[医药卫生—外科学]