转录调控基因GmLEC1的cDNA克隆及其植物表达载体的构建  

Molecular Cloning of Transcription Factor LEC1 and Construction of Plant Expression Vector for Sense and Anti-sense

作　　者：朱红林[1] 沙爱华[2] 符秀梅[1] 李小靖[1] 陈银华[1] 单志慧[2] 邱德珍[2] 周新安[2] 

机构地区：[1]海南大学教育部热带生物资源重点实验室,海南海口570228 [2]中国农业科学院油料作物研究所,湖北武汉430062

出　　处：《华北农学报》2009年第1期64-68,共5页Acta Agriculturae Boreali-Sinica

基　　金：中国农业科学院油料作物研究所基金;国家“863”计划(2006AA100104)

摘　　要：以高油大豆中豆32开花后30 d的种子为材料,根据已报道的拟南芥脂肪酸合成相关转录因子LEC1序列设计简并引物,采用同源序列法从大豆种子中分离了大小为850 bp的cDNA片段,测序结果表明,该片段与拟南芥中已克隆的脂肪酸合成基因高度同源,包含完整的读码框,采用酶切连接和gateway技术构建了该基因的超量表达和RNAi植物表达载体。为借助农杆菌介导法将LEC1基因转化到大豆再生植株中,对分离的LEC1基因进行功能验证,培育高油大豆新品种奠定了基础。The developing seeds of high oil soybean Zhongdou 32 were employed in this study. Degenerate primers were designed according to the reported sequence of transcription factor LECl related to fatty acid synthesis in Arabidopsis. A 850 bp cDNA fragment was isolated from soybean seed by homology-based candidate gene method. And RNAi vector pHGlec was constructed by gateway technology. The analysis of sequence revealed that the fragment was highly homologous with the cloned fatty acid synthesis gene in Arabidopsis, including complete reading frames. Over-expression vector was obtained by cleavage and ligation and RNAi vector pHGlec was constructed by gateway technology. This study laid a foundation for cultivating high oil soybean varieties by delivering LEC1 gene into soybean via Agrobacterium-mediated and functional verification of LEC1 gene.

关 键 词：LEC1基因 基因克隆 载体构建 大豆 

分 类 号：S565[农业科学—作物学] Q785[生物学—分子生物学]