高低含油量大豆成熟期胚抑制消减 cDNA文库构建及其差异表达基因分析

利用抑制性消减杂交（suppression subtractive hybridization, SSH）技术,以高含油量大豆品种“中豆32”和低含油量大豆品种“油春02-6”36天胚为实验材料构建消减cDNA文库。文库的插入片段集中在500bp左右，挑取766个克隆进行PCR筛选获得700个阳性克隆，经过点杂交筛选后选择了575个阳性克隆进行测序。经过同源性比对归并后得到237个差异表达基因，除10个未知基因外，其余涉及到蛋白贮藏、蛋白质降解、激素调节等多个方面。本文还对部分可能与油脂合成相关的差异表达基因进行了RT-PCR半定量和荧光定量PCR检测，并简要讨论了它们的功能，为进一步研究这些基因在大豆油脂合成过程中的作用奠定了基础。

Construction of suppression subtractive cDNA library and analysis of differentially expressed genes during maturing period seed of high and low oil content soybean variety

The 36 day after fertilization(DAF) embryoes of high oil content variety Zhongdou 32 and low oil content variety Youchun 02-6 were used to construct suppression subtractive cDNA library.The inserted fragment sizes focus on 500 bp,and 766 clones were verified by PCR,among which 700 clones were used to perform dot blot hybridization.575 clones were obtained and sequenced.237 differentially expressed genes were obtained by homologous BLAST search.Besides 10 unknown function genes,the others related to protein storing,protein degradation,auxin regulation and so on.Parts of differentially expressed genes probably relating to oil synthesis were detected by RT-PCR and Real-time quantitative PCR,their functions were also primarily discussed,which will be a guidance for studying the functions of these genes during oil synthesis.