农杆菌渗入法介导的基因瞬时表达技术和应用.pdfVIP

分子植物育种，2009 年，第7 卷，第5 期，第1032-1039 页 Molecular Plant Breeding, 2009, Vol.7, No.5, 1032-1039 新思路、新技术、新方法 Novel Thinking Technology 农杆菌渗入法介导的基因瞬时表达技术及应用 1,2 2,3 1,2 1,2 2* 邱礽 陶刚 李奇科 邱又彬 刘作易 1 贵州大学生命科学院, 贵阳, 550025; 2 贵州省农业生物技术重点实验室, 贵阳, 550006; 3 贵州省植物保护研究所, 贵阳, 550006 * 通讯作者, liuzuoyi@ 摘 要 农杆菌渗入法是在植物中瞬时表达外源基因的一种方法，该方法通过在植物叶片上注射农杆菌，使 目的基因转入植物细胞中进行快速表达。本文综述了农杆菌渗入法的原理、技术流程、影响因素及其应用，其 应用主要涉及外源基因的表达、检测转基因的表达、启动子分析、转录后基因沉默、抑制子功能鉴定、细胞定位 以及基因相互作用等方面。随着技术的进一步完善，我们相信该技术将成为转基因育种领域的重要的手段。 关键词 农杆菌渗入法, 转基因, 瞬时表达 Transient Gene Expression Mediated by Agroinfiltration and its Application Qiu Reng 1,2 Tao Gang 2,3 Li Qike 1,2 Qiu Youbin 1,2 Liu Zuoyi 2* 1 College of Life Sciences, Guizhou University, Guiyang, 550025; 2 The Key Laboratory for Agricultural Biotechnology of Guizhou, Guiyang, 550006; 3 Guizhou Institute of Plant Protection, Guiyang, 550006 * Corresponding author, liuzuoyi@ DOI: 10.3969/mpb.007.001032 Abstract Agroinfiltration is recently developed novel transgenic technology that can transiently express exoge- nous gene in plant by infecting the leaves with Agrobacteria which would facilitate the target gene to be integrated in- to plant genome and to rapidly express in the plant cells. This paper elaborated the principle of this technology, tech- nique procedure, influencing factor, and its application. The technologies would be widely employed to express ex- ogenous genes, analyze promoter function, silence post-transcriptional gene, and interact the genes. And it also used for identifying functional inhibitor and detecting genetically modified crops, etc. With the increasing development of this technology, we would believe that it will become one of the most important tools for transgenic breeding. Keywords Agroinfiltration, Transgene, Transient expression 将外源基因导入受体植物内，是研究基因功能 植物组织培养过程(Kapila et al., 1997) ；第二，表达效 和获得遗传修饰有机体的主要手段。根癌农杆菌介 率高。当单链的T-DNA 进入植物细胞后，不但转化 导法是目前最常用的遗传转化方法之一，但大多数 到植物基因组中