植物表达载体pCAMBIA2300-35S-GUS-CaMVterm的构建及验证  

Construction and Verification of an Plant Expression Vector pCAMBIA2300-35S-GUS-CaMVterm

作　　者：巩元勇[1,2] 冯永坤[2] 倪万潮[1] 束红梅[1] 郭书巧[1] 刘来华[2] 

机构地区：[1]江苏省农业科学院经济作物研究所农业部长江下游棉花和油菜重点实验室,南京210014 [2]中国农业大学资源、环境及粮食安全中心教育部植物与土壤互作重点实验室,北京100193

出　　处：《中国生物工程杂志》2013年第3期86-91,共6页China Biotechnology

基　　金：国家自然科学基金(31070223);江苏省自然科学基金(BK2010465);江苏省博士后科研资助计划(1201031B);国家转基因植物新品种培育科技重大专项(2011ZX08005-001)资助项目

摘　　要：为拓宽pCAMBIA2300表达载体的使用范围,进一步方便植物基因工程科研工作者在构建表达载体时的需要,利用SacⅠ和XhoⅠ双酶切pJIT166载体获取"35S-GUS-CaMVterm"片段,将该片段插入到pCAMBIA2300表达载体的多克隆位点区,构建完成了pCAMBIA2300-35S-GUS-CaMVterm表达载体。所构建的表达载体通过农杆菌介导转化拟南芥,获得拟南芥转基因植株,转基因植株的GUS染色结果进一步验证了本文所构建表达载体的正确性和实用性。该表达载体的成功构建,方便了构建基因超表达载体,以及使得研究启动子的活性及GUS组织化学定位成为可能,还可以通过GUS染色对转基因植物进行鉴定,为植物基因工程科研工作提供了一个较好的备选植物表达载体。Inorder to broaden the scope of use of plant expression vector pCAMBIA2300 and convenient to use for plant genetic engineering scientists in process of construction of expression vector, this research used SacⅠand XhoⅠdouble-digested pJIT166 vector obtaining 35S-GUS-CaMVterm fragment, then inserted this fragment to multiple cloning sites of pCAMBIA2300 vector, constructed the final expression vector pCAMBIA2300-35S-GUS-CaMVterm. The constructed expression vector was mediated by A. tumefacien （strain GV3101） using floral-dip method transformed to Arabidopsis thaliana. The GUS staining result of transgenic plants was further validated the correctness and practicality of this constructed vector. The construction of this expression vector was convenient for constructing gene over-expression vector,made it possible for studying promoter activity and GUS histochemical localization, and can also be used to identify transgenic plants. This constructed expression vector provided a better alternative plant expression vector for plant genetic engineering work.

关 键 词：植物表达载体 pCAMBIA2300 pJIT166 

分 类 号：Q812[生物学—生物工程]