| 伯氏疟原虫抗原基因高效植物表达载体的构建 |
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| 引用本文： | 王华,谭光宏,黄风迎,黄用豪,周松林,赵焕阁,林映莹.伯氏疟原虫抗原基因高效植物表达载体的构建[J].中国热带医学,2010,10(3):261-262,280. |
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| 作者姓名： | 王华 谭光宏 黄风迎 黄用豪 周松林 赵焕阁 林映莹 |
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| 作者单位： | 海南医学院海南省热带病重点实验室,海南,海口,571101 |
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| 基金项目： | 海南省自然科学基金项目(30624); 海南省教育厅高校科学研究资助项目(Hjkj200746) |
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| 摘 要： | 目的构建果实特异启动子驱动的含伯氏疟原虫裂殖子表面蛋白PbMSP4/5(Plasmodium berghei merozoitesurface protein4/5)基因的植物表达载体,并进一步提高抗原基因的表达量和免疫原性,为研制有效的转基因植物疟疾疫苗打下基础。方法分别以提取的番茄和伯氏疟原虫Anka株的基因组DNA为模板,扩增番茄果实特异表达启动子E8的核心序列(约1.11Kb)及PbMSP4/5基因(704bp),并通过重叠区扩增基因拼接法(Gene splicing by overlap extension,SOE)PCR将二者拼接,拼接后的序列为EM,并合成霍乱毒素B亚基基因CTB,共同构建重组质粒pCAMBIA1302-EM-CTB,电击法转化根癌农杆菌GV3103。结果重组质粒经酶切鉴定证明已成功转化。结论实验成功构建了以番茄果实特异启动子驱动PbMSP4/5基因、以CTB为黏膜免疫佐剂的高效植物表达载体。
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| 关 键 词： | 疟疾 植物口服疫苗 E8 PbMSP4／5 CTB |
Construction of an effective plant expression vector containing Plasmodium berghei antigen gene with cholera toxin B as adjuvant |
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| WANG Hua,TAN Guang-hong,HUANG Feng-ying,HUANG Yong-hao,ZHOU Song-lin,ZHAO Huan-ge,LIN Ying-ying.Construction of an effective plant expression vector containing Plasmodium berghei antigen gene with cholera toxin B as adjuvant[J].China Tropical Medicine,2010,10(3):261-262,280. |
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| Authors: | WANG Hua TAN Guang-hong HUANG Feng-ying HUANG Yong-hao ZHOU Song-lin ZHAO Huan-ge LIN Ying-ying |
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| Institution: | WANG Hua,TAN Guang-hong, HUANG Feng-ying,et al. (Hainan Provincial Key Laboratory of Tropical Medicine, Hainan Medical College, Haikou 571101, Hainan, P. R. China ) |
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| Abstract: | Objective To construct an effective plant vector driven by a fruit specific promoter for the expression of PbMSP4/5 (Plasmodium berghei merozoite surface protein 4/5)gene,further improve the expression an construction d immunogenicity of exogenous gene in plant,and prepare for the development of an effective anti-malaria plant edible vaccine. Methods Firstly,genomic DNA of tomato and Plasmodium berghei Anka was extracted,then according to SOE PCR(gene splicing by overlap extension) the core sequence (about ... |
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| Keywords: | Malaria Plant edible vaccine E8 PbMSP4/5 CTB |
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