利用抑制消减杂交技术分离辣椒细胞质雄性不育育性恢复相关EST

 以辣椒(Capsicum annuum L. ) 细胞质雄性不育系23A、121A, 和其相应的近等基因恢复系23C、121C为试验材料, 利用抑制消减杂交( SSH) 技术成功构建了CMS恢复基因诱导表达的消减cDNA文库。结合高密度点阵膜杂交差异筛选, 获得了282个阳性克隆。通过测序, 除去重复序列共得到175个Unique ESTs。在GenBank上进行BLAST分析, 55个EST片段未找到对应的同源序列, 可能代表了新基因;120个EST片段找到了对应的同源序列, 包括103个已知功能基因和17个未知功能基因。按照MIPS功能分类法, 将其分为14个功能组, 涉及代谢、胁迫应答、蛋白活性、转录因子、信号转导等多方面的功能。

Isolation of Fertility Restoration-related ESTs in Pepper Cytoplasmic Male Sterility Lines Using SSH

To isolate fertility restoration-related genes in pepper cytopasmic male sterility (CMS) lines, a forward subtracted cDNA librarywas constructed using supp ression subtractive hybridization (SSH).The cDNA of pepper CMS line 23A and 121A were used as driver and near2isogenic restorer line 23C and 121C as tester. Genes differentially exp ressed were screened using dot blot hybridization. A total of 175 non-redundant expressed sequence tags (ESTs) were identified. The genesppotential function analysis showed that 103 of these genes were homologous to known genes, and 55 could be new genes. The ESTs were sorted into 14 functional categories according to the Munich Information Center for Protein Sequences (MIPS) method.