辣椒细胞质雄性不育系和保持系SSH-cDNA文库的构建

Construction of SSH-cDNA Library for Cytoplasm Male Sterility Lines and Maintainer Lines in Pepper

以辣椒细胞质雄性不育系A1、A5为驱动方(driver),以其相应的保持系B1、B5为测验方(tester),利用抑制差减杂交技术构建了1个辣椒细胞质雄性不育保持系表达基因的正向差减cDNA文库。从差减文库中筛选到189个阳性单克隆,经高密度点阵膜杂交筛选和测序分析,共获得了42条表达序列标签(ESTs)。BLAST比对分析后,得到了35条有比对结果的序列,其中包括26条已知功能基因序列和9条未知功能基因序列,已知基因功能多与基础物质代谢、胁迫应答、信号转导等方面有关;有7条ESTs没有比对结果,可能代表一些新基因。

A forward subtracted cDNA library contained expression genes of pepper cytoplasm male sterility maintainer lines,was constructed by using suppression subtractive hybridization technique. The pepper cytoplasm male sterile lines A1 and A5 were used as drivers,and the corresponding maintainer lines B1 and B5 as testers. Based on the 189 positive monoclones selected from the subtracted library,42 expressed sequence tags（ESTs） were identified by DIG Nonradioactive Nucleic Acid Labeling and Detection System as well as sequencing. The BLAST analysis showed that 35 ESTs had gene potential functions,including 26 ESTs that are homologous to known genes,and 9 ESTs without homology. Most of the homologous gene functions are related to the basic material metabolism,stress defense,signal transduction;the 7 ESTs without BLAST results might imply some new genes.