抑制性差减杂交(SSH)技术在分离植物差异表达基因中的应用  

The Application of Suppression Subtractive Hybridization (SSH) on Isolating Plant Different Genes

作　　者：黄鑫[1] 戴思兰[1] 孟丽[1] 郑国生[2] 

机构地区：[1]北京林业大学园林学院,北京100083 [2]莱阳农学院生命科学院,青岛266109

出　　处：《分子植物育种》2006年第5期735-746,共12页Molecular Plant Breeding

基　　金：山东省自然科学基金项目“低温打破牡丹休眠机理的研究(Z2005D04)”资助.

摘　　要：抑制性差减杂交技术(SSH)是基于抑制PCR作用的cDNA差减杂交,是在转录水平上研究差异基因表达的技术。该技术在一个循环过程中完成差异表达基因丰度的均等化及目标群体和对照群体中相同基因的去除,从而实现差异表达基因的高度富集。一个典型的SSH过程,可在一个差减杂交过程中将稀有基因序列富集上千倍。本文通过详细分析该技术在植物差异表达基因分离中的应用发现:SSH技术主要用于分离组织特异性表达和诱导型表达的基因。首先,SSH技术广泛应用在分离植物发育过程中不同发育阶段和不同组织器官中的组织特异性表达的基因,为揭示植物生长发育过程的分子机理提供了有效手段;另外,在不同植物中,该技术被大量应用于分离各种生物及非生物胁迫条件下诱导表达的抗性相关基因,可以揭示植物抗逆的分子机理。目前,SSH技术已开始应用于分离与次生代谢产物合成相关的基因。SSH技术是分离植物差异表达基因,揭示植物复杂生命现象分子机理的有效方法,将在植物差异表达基因研究中得到更广泛的应用。Suppression subtractive hybridization （SSH） is a technique for the study on different genes expression on transcriptional level, which is a cDNA subtractive hybridization based on the suppression PCR. It combines normalization and subtraction in a single procedure. The normalization step equalizes the abundance of cDNAs within the target population and the subtraction step excludes the common sequences between the target and driver populations. In a model system, SSH technique amplify rare sequences over 1 000 times in one round of subtractive hybridization. The application of SSH on isolating plant differential expression genes were reviewed in this paper. The authors proposed that SSH was mainly used to isolate tissue specific genes and inducible expression genes. Firstly, the SSH was used to isolate tissues or organs specific expression genes in different development stages of plant. These genes provide evidence for explaining the molecular mechanism of plant development. Then SSH was widely used to isolate genes related to resistance under biological or abiological stress in different plants and the molecular mechanism of the resistance was primarily elucidated. And now, the technique was applied to isolate genes related to biosynthesis of second products. SSH is an effective measure to isolate differential expression genes and illuminate the complex phenomena in plant life.

关 键 词：抑制性差减杂交(SSH) 植物 差异表达基因 

分 类 号：Q943.2[生物学—植物学]