| 高山离子芥冷诱导基因转化甘蔗二元植物表达载体构建 |
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| 引用本文： | 卢双楠,李粲,滕峥,刘开雨,刘芳,邱永福,梁朝旭,方位宽,何姗珊,刘晓静,李鸣,梁俊,李容柏.高山离子芥冷诱导基因转化甘蔗二元植物表达载体构建[J].南方农业学报,2012,43(9):1262-1268. |
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| 作者姓名： | 卢双楠 李粲 滕峥 刘开雨 刘芳 邱永福 梁朝旭 方位宽 何姗珊 刘晓静 李鸣 梁俊 李容柏 |
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| 作者单位： | 广西大学生命科学与技术学院,南宁530005广西作物遗传改良重点开放实验室,南宁530007广西大学农学院,南宁,530005广西甘蔗遗传改良重点实验室/广西农业科学院甘蔗研究所,南宁,530007广西甘蔗遗传改良重点实验室/广西农业科学院甘蔗研究所,南宁530007亚热带农业生物资源保护利用重点实验室,南宁,530005 |
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| 基金项目： | 广西科学基金项目(桂科自0990182,桂科基0778006-4,桂科青0832059,2011GXNSFF018002,2011GXNSFD018021);广西农业科学院博士后基金项目(桂农科博2009013);广西农业科学院基本科研业务专项项目(201107Z基,G2010003,G2010003);广西农业科学院公益性维持项目(桂农科2012GW13);南宁市科学研究与技术开发计划项目(201102026B) |
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| 摘 要： | 目的]利用载体重组技术分别将高山离子芥冷诱导基因(Cbcor15a)和报告基因eGF插入载体pCambia1300-bar,并引入玉米泛素基因启动子UBi-1代替载体本身启动子CaMV 35s,重组为适合甘蔗转基因的二元植物表达载体pCambia 1300-cbcor 15a-bar.方法]参照pCambia1300-bar载体多克隆位点和基因Cbcor15a、eGFP和启动子UBi-1的核苷酸序列设计引物,通过载体重组技术将基因和启动子分别插入相应的位点.利用基因枪分别将pCambia1300-bar载体和重组载体导入洋葱表皮细胞,用荧光显微镜和激光共聚焦显微镜观察.结果]与导入pCambia 1300-bar载体的洋葱表皮细胞相比较,导入重组载体pCambia1300-cbcor15a-bar的洋葱表皮细胞内有强烈的绿色荧光信号.结论]重组甘蔗转基因二元植物表达载体启动子Ubi-1能够调控下游冷诱导基因Cbcor15a和报告基因eGFP的正常高效表达,为外源基因Cbcor15a转化甘蔗提供保障.
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| 关 键 词： | Cbcor15a 甘蔗 转基因 植物表达载体构建 瞬时表达 |
Construction of bivalent plant expression vector with cold-induced gene of Chorispora bungeana in sugarcane |
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| LU Shuang-nan,LI Can,TENG Zheng,LIU Kai-yu,LIU Fang,QIU Yong-fu,LIANG Zhao-xu,FANG Wei-kuan,HE Shan-shan,LIU Xiao-jing,LI Ming,LIANG Jun,LI Rong-bai.Construction of bivalent plant expression vector with cold-induced gene of Chorispora bungeana in sugarcane[J].Journal of Southern Agriculture,2012,43(9):1262-1268. |
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| Authors: | LU Shuang-nan LI Can TENG Zheng LIU Kai-yu LIU Fang QIU Yong-fu LIANG Zhao-xu FANG Wei-kuan HE Shan-shan LIU Xiao-jing LI Ming LIANG Jun LI Rong-bai |
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| Institution: | 5 *(1College of Life Science and Technology,Guangxi University,Nanning 530005,China;2 Guangxi Crop Genetic Improvement and Biotechnology Laboratory,Nanning 530007,China;3 Agricultural College,Guangxi University,Nanning 530005,China;4 Guangxi Key Laboratory for Sugarcane Genetic Improvement /Sugarcane Research Institute,Guangxi Academy of Agricultural Sciences,Nanning 530007,China;5 State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,Nanning 530005,China) |
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| Abstract: | Objective]A new bivalent plant expression vector,named pCambia1300-cbcor15a-bar,was recombined by inserting two genes,i.e.Cbcor15a and eGFP,into the vector pCambia1300-bar and replacing the promoter CaMV 35s with Ubi-1.Method]Based on the multiple cloning sites of the expression vector pCambia1300-bar,the primers were designed according to the nucleotide sequence of gene Cbcor15a and eGFP and the promoter Ubi-1,and then the fragments of the genes and promoter were inserted into the vector pCambia1300-bar.The plasmids of the vector pCambia1300-cbcor15a-bar were transduced into onion epidermal cells via particle bombardment.The results were observed through fluorescence microscopy and confocal laser scanning microscope,respectively.Result]Compared to onion epidermal cell with pCambia1300-bar,onion epidermal cell transduced with the recombinant vector plasmid had a very bright green florescence.Conclusion]Downstream cold-induced gene Cbcor15a and reporter gene eGFP regulated by up-stream promoter Ubi-1 were expressed efficiently,which could ensure the construction of Cbcor15a. |
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| Keywords: | Cbcor15a sugarcane transgene construction of plant expression vector transient expression |
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