油菜MADS-box家族基因AGL13表达分析及RNA干扰载体的构建  

Expression and construction of RNA interference vector of MADS-box family gene AGL13 from Brassica napus

作　　者：熊书[1] 孙厚良[1] 杨丽珊[1] 马强[1] 李国利[1] 周大祥[2] XIONG Shu;SUN Hou-liang;YANG Li-shan;MA Qiang;LI Guo-li;ZHOU Da-xiang(Department of Basic Medicine,Chongqing Three Gorges Medical College,Wanzhou 404120;College of Life Science and Engineering,Chongqing Three Gorges University,Wanzhou 404100,China)

机构地区：[1]重庆三峡医药高等专科学校基础医学部,万州404120 [2]重庆三峡学院生命科学与工程学院,万州404100

出　　处：《生物学杂志》2018年第5期45-48,共4页Journal of Biology

基　　金：重庆市自然科学基金(cstc2016jcyjA2132);重庆市教委科学技术研究项目(KJ1502601)

摘　　要：为探索油菜种子中油脂合成的分子调控机理,提高油脂含量,改良油菜品质奠定基础。通过定量PCR分析油菜AGL13基因的组织表达模式,同时构建AGL13的干扰载体。生物信息学分析发现油菜AGL13具有完整且典型的MADS-box保守结构域,蛋白整体形态呈现典型的DNA核酸结合结构,属于MADS-box家族转录调控因子,亲源关系与拟南芥AGL13蛋白最近。定量PCR研究发现AGL13在油菜盛花期到种子45DAP(盛花后45 d的种子)表达水平都很高,其中在果夹15DAP表达水平最高,实验成功构建了RNA干扰载体。结果表明,AGL13基因在油菜种子发育和油脂积累过程中强烈表达,该基因可能与油菜种子发育及油脂的合成密切相关。RNA干扰载体的构建成功将为后续获得转基因油菜,观察转基因油菜胚的发育和种子油脂的积累,从而明确AGL13在油菜胚的发育及油脂的合成中发挥着重要作用。In this study, the tissue expression pattern of AGL13 was analyzed by qPCR, and the RNA interference vector was constructed to explore the molecular regulation mechanism of oil synthesis in seed of Brassica napus to increase the oil content. Bioinformatics analysis showed that AGL13 protein of Brossica napus has complete and typical MADS-box conserved domain. The overall shape of protein showed typical DNA nucleic acid binding structure. The AGL13 belonged to the MADS-box family transcription factor, and genetic relationship was the closest with AGL13 protein of Arabidopsis . The qPCR study found that AGL13 expression was very high from the flowering stage to the seed 45DAP （ays seed after 45 d flowering）, was highest in the fruit clamp 15DAP. The RNA interference vector was successfully constructed in the experiment. The results showed that the AGL13 gene was strongly expressed in seed development and oil accumulation, which may play an important role in the seed development and oil synthesis. The successful construction of RNA interference vector would be favourable for obtaining the transgenic B. napus , and observing the seed development and oil accumulation of transgenic B. napus . Thus determining the AGL13 gene plays an important role in the seed development and oil synthesis.

关 键 词：油菜 MADS-BOX AGL13 RNAI 油脂合成 

分 类 号：Q786[生物学—分子生物学] Q782