玉米温敏自交不亲和材料花丝SSH-cDNA文库的构建

Construction of SSH-cDNA library for silks of a thermo-sensitive genic self-incompatible line in maize

为了解玉米温敏自交不亲和材料HE97的表达机制,构建了该材料自交亲和与不亲和阶段花丝的正向与反向抑制差减杂交文库,并对其中的219个正向克隆和52个反向克隆进行了测序分析,分别获得了49个和20个Unique ESTs,涉及到代谢、光合作用、信号转导、物质运输、蛋白质合成加工、防御反应、细胞的发育和衰老、基因表达等重要生命过程中的12种不同功能类型.通过电子克隆获得了包含1 090 bp的烯脂酰辅酶A水合酶基因,并对其蛋白质功能进行了分析.

To understand the expression mechanism of a thermo-sensitive genic self-incompatible line HE97 in maize, the forward and reverse library was constructed by means of suppression subtractive hybridization （SSH） method, using the different silks as the material,taken from the self-compatible and the self-incompatible stages. After 219 forward clones and 52 revise clones were sequenced, 49 unique expressed sequence tads （ESTs） in forward library and 20 unique ESTs in reverse library were obtained by clustering and blast analysis. The ESTs were sorted into 12 different functional categories, involving in the process of metabolism, transcription, signal transduction, cell growth/division, transporters, and diseases defence. One enoyl CoA hydratase gene with 1 090 bp was cloned by silicon cloning, and the protein function was analyzed.