Sternberg Michal Bassani, Gepstein Shimon
Faculty of Biology, Technion, Isreal Institute of Technology, Haifa, Isreal.
Methods Mol Biol. 2007;371:289-305. doi: 10.1007/978-1-59745-361-5_21.
Molecular and cellular processes related to the senescence syndrome are determined by programs of differential gene expression. Subtractive cDNA hybridization is a powerful approach to identify and isolate differentially expressed genes in various systems. A highly effective method, termed suppression subtractive hybridization (SSH), has been applied for the generation of subtracted cDNA library of senescing leaves. The method consists of two main stages, the normalization step that equalizes the abundance of cDNAs within the target population and the subtraction step that eliminates the common sequences between the target and the driver populations. The successful generation of library containing high numbered of rare and abundant cDNA clones in senescing plant cells proves the applicability of this method for global identification of differentially expressed genes during cellular senescence.
与衰老综合征相关的分子和细胞过程由差异基因表达程序决定。消减cDNA杂交是在各种系统中鉴定和分离差异表达基因的有力方法。一种高效的方法,称为抑制消减杂交(SSH),已被应用于生成衰老叶片的消减cDNA文库。该方法包括两个主要阶段,即使目标群体内cDNA丰度均等化的标准化步骤,以及消除目标群体和驱动群体之间共同序列的消减步骤。成功构建出包含大量衰老植物细胞中稀有和丰富cDNA克隆的文库,证明了该方法在细胞衰老过程中全面鉴定差异表达基因的适用性。