桑树肌动蛋白MaACT3启动子的克隆及农杆菌介导的瞬时表达  

Cloning of MaACT3 Promoter from Mulberry (Morus alba) and Agrobacterium-Mediated Transient Expression in Leaves

作　　者：李军[1] 赵爱春[1] 王茜龄[1] 祝娟娟[1] 张琼予[1] 金筱耘[1] 李镇刚[1] 鲁成[1] 吴存容[1] 余茂德[1] 

机构地区：[1]西南大学生物技术学院/家蚕基因组生物学国家重点实验室,重庆400715

出　　处：《中国农业科学》2012年第4期625-632,共8页Scientia Agricultura Sinica

基　　金：国家现代农业产业技术体系建设专项(nycytx-27-gw101);国家自然青年基金(31101769);重庆市蚕桑重大科技专项(CSTC;2009AA1024)

摘　　要：【目的】克隆桑树肌动蛋白MaACT3的启动子。【方法】采用抑制PCR技术获得肌动蛋白的5′端侧翼序列;采用农杆菌感染成熟叶片瞬时表达检测启动子活性。【结果】获得的5′端侧翼序列含有基因的翻译起始位点、第一外显子、第一内含子和第二外显子的一部分,外显子部分与已报道的桑树肌动蛋白MaACT3序列完全相同。通过荧光显微镜观察到了绿色荧光,在mRNA水平上也检测到表达,证实了获得的5′端序列具有启动活性。【结论】获得的桑树肌动蛋白MaACT3的启动子具有启动活性,可以应用于桑树转基因研究。【Objective】 The objective of this study is to clone the promoter of MaACT3 from mulberry（Morus alba）.【Method】The promoter of MaACT3 was obtained by suppression PCR method and the activity of promoter was valued by agrobacterium-mediated transient expression.【Result】 The sequence analysis indicated that exon I,intron I and partial exon II were included and the partial exon II was the same as mRNA of MaACT3 reported before.Mature leaves were used for agrobacterium-mediated transient expression.Green fluorescence was measured by fluorescence microscope and mRNA was also detected.【Conclusion】 The promoter of MaACT3 from mulberry expressed promoter activity and could be used in transgenic research of mulberry.

关 键 词：桑树 接头 启动子 绿色荧光蛋白 

分 类 号：S888[农业科学—特种经济动物饲养]