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甜瓜果实酸性转化酶基因cDNA片段的克隆、反义表达载体的构建和烟草的遗传转化
Cloning of a cDNA Fragment of Acid Invertase Gene from Cucumis Melo L. Fruit, constructing of Antisense expression vector and transferring of tobacco
【Author】 YU Xi-Yan,ZHAO Shuang-Yi,HE Qi-Wei(Shool of Horticulture,Agriculture University,Taian 270118) (Shoolof Life Sciences,Shandong University,Jinan250100) (Vegetable Jnstitute,Shandong Academy of Agriculture Sciences Jinan 250100)
【机构】 山东农业大学园艺学院； 山东大学生命科学院； 山东省农科院蔬菜所；
【摘要】 采用RT-PCR方法,从甜瓜果实总RNA中扩增出目标cDNA片段(MAI1),在GenBank中登记号为AF490425.将该基因片段定向插入到pROK2的BamH Ⅰ/Sma Ⅰ克隆位点,构建了Anti-MAI1植物表达双元载体,在根癌农杆菌介导下转化烟草,经PCR和PCR-Southern杂交检测,证明此基因已整合入烟草的核基因组中.
【Abstract】 The aimed cDNA fragment (MAI1) was amplified from the total RNA isolated from melon fruit via RT-PCR. Its access number in GenBank is AF490425.This fragment was inserted between BamH I and Sma I of pROK2 for constructing Anti-MAI1 binary vector and transferring it into tobacco. PCR and PCR-Southern proved that MAI1 had been conformiting into the genomic DNA of tobacco.
【Key words】 Melon fruit； RT-PCR； Acid invertase gene； Antisense vector； Tobacco； Transformation；
- 【会议录名称】 蔬菜分子育种研讨会论文集
- 【会议名称】蔬菜分子育种研讨会
- 【会议时间】2004-06
- 【会议地点】中国北京
- 【分类号】S652
- 【主办单位】中国园艺学会蔬菜专业委员会、中国农业科学院蔬菜花卉研究所