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梨韧皮部特异表达启动子AtSUC2驱动下的GUS基因的转化和表达
引用本文:孙清荣,孙洪雁,赵衍,茹斯·海门得,罗伯特·戴维斯.梨韧皮部特异表达启动子AtSUC2驱动下的GUS基因的转化和表达[J].园艺学报,2008,35(4):487-492.
作者姓名:孙清荣  孙洪雁  赵衍  茹斯·海门得  罗伯特·戴维斯
作者单位:( 山东省果树研究所,山东泰安271000; 美国农业部百思维尔农业研究中心,植物分子病理实验室,马里兰20705,美国)
基金项目:山东省优秀中青年科学家科研奖励基金
摘    要: 以 ‘Old Home’ 梨无菌苗叶片为外植体, 利用根癌农杆菌介导将专一在韧皮部表达启动子AtSUC2 驱动下的GUS 基因转入 ‘Old Home’中。筛选出了‘Old Home’ 梨叶片高效遗传转化的最佳条件:农杆菌和外植体在液体培养基上共培比在固体培养基上共培转化率提高5倍。 PCR分析和Southern 杂交鉴定表明GUS 基因已整合到转基因植株的基因组中,组织化学检测和Western 杂交鉴定证明了GUS 基因在转基因植株韧皮部中的表达。

关 键 词:  启动子  韧皮部特异表达  基因转化
收稿时间:2007-12-27

Transgene Expression in Pear (Pyrus communis L. ) Driven by Phloem-specific Promoter AtSUC2
SUN Qing-rong,SUN Hong-yan,ZHAO Yan,Rosemarie Hammond,Robert E.Davis.Transgene Expression in Pear (Pyrus communis L. ) Driven by Phloem-specific Promoter AtSUC2[J].Acta Horticulturae Sinica,2008,35(4):487-492.
Authors:SUN Qing-rong  SUN Hong-yan  ZHAO Yan  Rosemarie Hammond  Robert EDavis
Institution:(Shandong Institute of Pomology, Tai'an, Shandong 271000, China; Molecular Plant Pathology Laboratory, USDA-Agriculture Research Service, Beltsville, MD 20705, U.S.A.)
Abstract:Leaves of 'Old Home' Pear (Pyrus communis L.) in vitro were selected as explants. GUS (β-glucuronidase) reporter gene (uidA) under the control of the Arabidopsis sucrose-H+ symporter gene (AtSUC2) promoter which is phloem-specific was introduced into 'Old Home' by Agrobacterium tumefaciens. Optimal conditions were established for efficient transformation from leaf explants of a pear cultivar, 'Old Home'. High transformation efficiency was achieved by an improved induction stage following initial Agrobacterium infection. In the induction stage, Agrobacterium cells and parent leaf explants were co-cultivated on a liquid induction medium, which yielded a five-fold increase of transformation frequency over conventional co-cultivation on a solid medium. The integration was checked by PCR and Southern hybridization. The expression of uidA gene was analyzed by GUS histochemical detection and Western blot analysis.
Keywords:pear  promoter  phloem-specific expression  transgene
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