mRNA差异显示技术研究进展  

Progress in the Technique of mRNA Differential Display

作　　者：李文雍[1] 陈清轩[1] 

机构地区：[1]中国科学院遗传与发育生物学研究所

出　　处：《农业生物技术学报》2004年第5期597-601,共5页Journal of Agricultural Biotechnology

基　　金：国家重大基础研究发展规划(973)项目(No.G2000016107)资助

摘　　要：分离和鉴定差异表达基因是分子生物学研究领域的热点之一。DDRT-PCR 是目前该领域中最受青睐的技术。它有简便、灵敏和 RNA 用量少、且可同时比较多组样品等优点,但也存在假阳性率高、所得的差异片段较短等不足。针对这些缺陷。人们在引物的设计、DDRT 和 PCR 参数的优化、差异显示凝胶的选择、阳性克隆的鉴定等环节进行了改进,并与其它生物技术结合,发展了该技术,如代表性差异分析、荧光 mRNA 差异显示和单个植入前胚胎 mRNA 差异显示技术,使该技术日臻完善。结合本研究室近年来的研究工作,对 DDRT-PCR 技术的原理、优越性、主要缺陷、技术改进及应用前景等方面进行简要综述。It is one of the most importance technigues in molecular biology study to identify and analyze differential expressed genes between control cell and experimental cell or tissues.Differential display reverse transcription-polymerase chain reaction (DDRT-PCR)technique is preferred in this research field currently for it owns some strongn points such as simple,sensitive,few quantity of RNA,and can display differences from several samples.But it also has shortcomings,for example the high rate of fake positive clones,the shorter of differential fragments obtained.To overcome these limitations,several taches including total RNA, primers,the parameters of DDRT and PCR,selection of sequencing gel and identification of clones have been modified.And it has been improved to be perfectly match other biological approaches.Such as represent difference analysis(RDA),fluorescein differential display(FDD),and single preimplantation embryo differential display reverse transcription PCR(SPEDDRT-PCR).The theory, advantages,main limitations and modifications of this method have been reported in this paper combining our research work.

关 键 词：MRNA差异显示技术 DDRT-PCR 研究进展 差异表达基因 假阳性率 鉴定 分子生物学研究 植入前胚胎 阳性克隆 代表性差异分析 

分 类 号：S511[农业科学—作物学] Q786[生物学—分子生物学]