植物源内含子对GUS基因表达模式的影响  

Effects of Botanical Intron on Expression Pattern of GUS Gene

作　　者：邹智[1] 吴刚[1] 肖玲[1] 武玉花[1] 聂淑晶[1] 肖娜[1] 卢长明[1] 

机构地区：[1]中国农业科学院油料作物研究所农业部油料作物遗传改良重点开放实验室,武汉430062

出　　处：《生物技术通报》2008年第6期78-82,共5页Biotechnology Bulletin

基　　金：国家自然科学基金项目(30471009)

摘　　要：采用报告基因的瞬间表达来优化转化体系是提高农杆菌介导法转化效率的重要手段,GUS以稳定、易于定性定量分析等独特优点成为瞬间表达体系的首选。pCaMV 35S启动下的GUS基因可在农杆菌中表达,另外还首次报道该嵌合基因同时也可在大肠杆菌中高效表达。为避免GUS基因在农杆菌中的表达对瞬间表达体系的影响,一拟南芥蛋白基因的内含子被插入到GUS基因的编码区,进而构建了含内含子的GUS植物表达载体pBI121-GUSint。组织化学染色结果表明,GUSint在农杆菌中没有表达,而在接种3d的油菜中可高效瞬间表达,其中在子叶柄(带子叶)中瞬间表达率高达100%,这一方面证实载体构建成功,另一方面也为进一步优化油菜及其它芸薹属植物转化体系及在油菜中快速研究目的基因的功能和表达调控模式奠定了基础。Using transient expression of reporter genes to optimize transformation systems has become an important method to improve the efficiency of Agrobacterium-mediated transformation,while GUS gene used as reporter gene commonly in transient expression systems to analyze transformation qualitatively and quantitatively.GUS gene drived by pCaMV 35S was expressed in Agrobacterium tumefaciens,and expression in E.coli was reported for the first time.To avoid GUS expressing in Agrobacterium,a GUS expression vector pBI121-GUSint with a histone intron of Arabidopsis thaliana was constructed.The results of histochemical GUS assay showed that no expression of GUSint was observed in Agrobacterium,with transient expression as high as 100% in cotyledonary node（with cotyledons） of Brassica napus L.after inoculated for three days.It not only verified that the vector was successfully constructed,but also laid a basis for further increasing the stable transformation efficiency of other Brassica plants,and rapidly clarifying the function and pattern of expression and regulation of a unknown gene.

关 键 词：GUS 内含子 表达模式 根癌农杆菌 甘蓝型油菜 瞬间表达 

分 类 号：Q943[生物学—植物学]