拟南芥植物型PEPC基因人工小RNA表达载体的构建与遗传转化  

Construction of Plant-type Phosphoenolpyruate Carboxylase (PEPC) Gene Artificial miRNA Expression Vector and Transformation in Arabidopsis thaliana

作　　者：冯都华[1,2] 王伏林[2] 陈锦清[2] 

机构地区：[1]浙江师范大学化学与生命科学学院,浙江金华321004 [2]浙江省农业科学院病毒学与生物技术研究所,杭州310021

出　　处：《中国农学通报》2011年第12期249-254,共6页Chinese Agricultural Science Bulletin

基　　金：国家自然科学基金项目"油料作物高油种质资源创制的机理及其应用"(30430450);"油菜籽脂肪酸生物合成的双重遗传调控"(30370910)

摘　　要：磷酸烯醇式丙酮酸羧化酶(PEPC)是植物中具有多种生理功能的酶。为探索拟南芥中植物型PEPC基因对模式植物拟南芥脂肪酸含量以及抗逆性等方面的影响,笔者构建了同时敲除拟南芥Atppc1、Atppc2和Atppc3基因的人工小RNA(amiRNA)植物表达载体pFGC-amiAtppc123,经根癌农杆菌EHA105介导,用花序浸染法转化拟南芥,成功获得转基因植株。RT-PCR半定量分析表明人工小RNA在转化植株中成功进行了超量表达。该试验为分析拟南芥脂肪酸含量以及抗逆性方面提供了基础材料。To investigate the functions of PEPC in lipid content,fatty acid composition and the response to environmental stress in C 3 plants,Arabidopsis thaliana plant-type PEPC genes（referred to as Atppc1,Atppc2 and Atppc3） were silence by the artificial miRNA（amiRNA） technique.The author constructed the plant expression vector named pFGC-amiAtppc123 and transformed it into Arabidopsis thaliana via in floral dip method.Transgenic plants were confirmed by PCR.The amiRNA transcript levels were detected by semi-quantitative RT-PCR.The results showed that amiRNA expression was drastically increased in transgenic plant compared with non-transgenic plants.

关 键 词：PEPC 拟南芥 amiRNA 脂肪酸含量 花序浸染 

分 类 号：Q78[生物学—分子生物学]