| 玉米特异启动子驱动下结核Hsp65与Esat-6融合基因表达载体的构建及鉴定 |
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| 引用本文： | 李君武,王珊珊,宋东,刘艳,黄清华. 玉米特异启动子驱动下结核Hsp65与Esat-6融合基因表达载体的构建及鉴定[J]. 华北农学报, 2009, 24(3) |
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| 作者姓名： | 李君武 王珊珊 宋东 刘艳 黄清华 |
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| 作者单位： | 暨南大学医学院,微生物与免疫学教研室,广东广州,510632 |
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| 基金项目： | 广东省科技计划重大专项，广东省科技计划重点项目 |
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| 摘 要： | 利用转基因玉米研制新型结核口服疫苗.构建植物双元表达质粒pCAMBIA1300GHLE,并转化农杆菌LBA4404.以本实验室构建的pEGHLE为模板经聚合酶链反应(PCR)扩增出Hsp65-Esat6基因,连接到含有玉米特异性启动子globulin-1的pCR2.1载体上;将globulin1-HLE联合片断切下连到含有抗除草剂基因bar的pCAMBIA1300载体中;电击法将重组质粒转化到农杆菌LBA4404中.成功构建了pC1300GHLE质粒,酶切鉴定得到3.3 kb和8.6Kb2条带,测序分析表明克隆的H8p65和Esat6序列与NCBI上公布序列一致;成功转化到农杆菌中,酶切从农杆菌中所提的质粒,条带大小与预期结果相符合.成功构建和转化了pCl300GHLE表达载体,为成功研制利用转基因植物生产抗结核口服疫苗奠定了基础.
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| 关 键 词： | 结核分支杆菌 热休克蛋白65 |
Construction of Vector for Expression of Hsp65 and Esat-6 Genes Driven by a Maize Endosperm-specific Promoter |
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| LI Jun-wu,WANG Shan-shan,SONG Dong,LIU Yan,HUANG Qing-hua. Construction of Vector for Expression of Hsp65 and Esat-6 Genes Driven by a Maize Endosperm-specific Promoter[J]. Acta Agriculturae Boreali-Sinica, 2009, 24(3) |
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| Authors: | LI Jun-wu WANG Shan-shan SONG Dong LIU Yan HUANG Qing-hua |
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| Affiliation: | Department of Microbiology and Immunology;Medical College of Jinan University;Guangzhou 510632;China |
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| Abstract: | To construct the plant expression plasmid containing Mycobacterium tuberculosis Hsp65 and Esat-6 genes,and transform the recombined vector into Agrobacterium tumerfaciens LBA4404.The fusion DNA fragment of Hsp65 and Esat-6 were amplied from pEGHLE by polymerase chain reaction(PCR)were cloned into the vector pCRG.The combine fragment of promoter globulin-1 and the target gene HLE which get from doubled enzymes digestion of the recombined plasmid pCRGHLE was inserted into the plant expression vector pCAMBIA13... |
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| Keywords: | Esat-6 globulin-1 |
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