杨树木质部特异启动子5'端侧翼序列缺失表达载体构建及转化烟草的研究  

Construction of Deletion Expression Vectors for Poplar Xylem Specific Promoter 5' Flanking Sequence and Its Transformation into Tobacco

作　　者：赵金风[1] 李杰[1] 张爽[1,2] 杜克久[1,2] 

机构地区：[1]河北农业大学,河北保定071000 [2]河北省林木种质资源与森林保护重点实验室,河北保定071000

出　　处：《蚕业科学》2013年第5期858-861,共4页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(No.31000305);河北省自然科学基金项目(No.C2010000683)

摘　　要：采用5'端系列缺失分析法,PCR扩增杨树木质部特异性启动子JCesAP中与转录起始位点距离不同的5'端缺失启动子片段ZU2、ZU3、ZU4。为鉴定缺失片段的功能活性,以β-葡糖醛酸酶基因(GUS)作为报告基因,分别构建JCesAP启动子3个缺失片段的表达载体,并通过农杆菌介导瞬时转化烟草。对转化烟草植株进行GUS活性检测的结果表明,缺失片段ZU3、ZU4有启动子活性,且ZU4启动子活性强度与CaMV35S启动子相当,转化烟草的叶片表面都呈现大面积蓝色,而ZU2没有启动子活性。木质部特异性核心启动子的筛选将有助于利用转基因技术治理桑天牛等天牛类蛀干害虫。Using 5＇ end serial deletion analysis method, xylem-specific promoter JCesAP5＇ deletion promoter fragments ZU2, ZU3 and ZU4 which have different distances with transcription start site were amplified by PCR. To detect functional activity of the deletion fragments, expression vectors carrying these 3 JCesAP promoter deletion fragments were con- structed and transiently transformed into tobacco via Agrobacterium mediation with β-glucuronidase gene （GUS） as the reporter gene. Determination of GUS activity in transformed tobacco plants revealed that ZU3 and ZU4 had promoter ac- tivity, and ZU2 had no promoter activity. Moreover, the promoter activity of ZU4 was close to that of promoter CaMV35S. ZU4 transformed tobacco plant had large blue areas on its leaf surface. Screening of xylem-specific core promoters will facilitate the control of trunk borers of Cerambycidae including Apriona germari by using transgenic technique.

关 键 词：杨树 木质部特异启动子 缺失分析法 载体构建 瞬时表达 

分 类 号：S792.11[农业科学—林木遗传育种]