绥农14鼓粒期籽粒cDNA文库构建及初步分析  

Construction and characterization of the filling stage's seed cDNA library from Suinong14( Glycine max)

作　　者：王跃平[1,2] 李英慧[2] 陈雄庭[1] 常汝镇[2] 邱丽娟[2] 

机构地区：[1]中国热带农业科学院热带作物生物技术国家重点实验室/热带生物技术研究所,海口571101 [2]中国农业科学院作物科学研究所,国家农作物基因资源与遗传改良重大科学工程,农业部作物种质资源与生物技术重点开放实验室,北京100081

出　　处：《中国油料作物学报》2008年第1期40-45,共6页Chinese Journal of Oil Crop Sciences

基　　金：国家“863”课题(2006AA10A110);国家自然科学基金重大项目(30490251)

摘　　要：大豆EST分析对深入了解大豆重要性状形成机制并发掘新基因具有重要意义。本研究利用SMART技术,构建了绥农14鼓粒期的籽粒cDNA文库并测定EST序列,目的是分析绥农14鼓粒期的基因表达特性,为挖掘和克隆基因提供物质支撑。结果表明,本研究构建文库的初始库容为1.2×106,扩增后文库的库容为1.6×109,重组率达98%,用载体两端通用引物进行PCR鉴定,插入片段均大于0.9kb,随机对2300个单克隆测序,获得有效序列2201条,SeqMANⅡ组装整理后的序列2114条,包括1516个contig或者singlet,平均长度为672bp。对组装后的序列构建籽粒鼓粒表达图谱,发现籽粒形成初期,能量代谢、初级代谢途径、蛋白合成定向贮藏类基因表达量较多,而涉及细胞运输,细胞结构和抗性类基因表达量较低。Understanding the genes related to soybean diseases and characteristics will increase the crop yield and quality by Expressed Sequence Tags （EST） analysis. We constructed the cDNA library of Suinong 14 in the filling-time seed by SMART Kit. The library＇s content was 1.2×106, the recombination ratio was nearly 98% and the average inserts ware more than 0.9 kb. Use the SeqMan II software to assembe 2 114 EST sequences which included 1 516 unigenes, and the average length was 672bp. After construct the EST expression profile in the very young seed time, we found the primary metabolism, energy metabolism and protein synthesis classes genes were highly expressive. And the cell traffic, cell organization and defense genes were mostly restricted.

关 键 词：CDNA文库 表达序列标签(EST) 测序 同源分析 功能注释 表达图谱 

分 类 号：S565.103[农业科学—作物学]