| 小鼠CD86基因启动子的构建及活性分析 |
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| 引用本文： | 范敏其,龚卫娟,龚春香,肖炜明,丁岩冰,季明春. 小鼠CD86基因启动子的构建及活性分析[J]. 现代免疫学, 2009, 0(5) |
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| 作者姓名： | 范敏其 龚卫娟 龚春香 肖炜明 丁岩冰 季明春 |
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| 作者单位： | 扬州大学动物科学与技术学院繁殖教研室;扬州大学医学院免疫学教研室;扬州大学第二附属医院消化病研究室; |
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| 基金项目： | 国家自然科学基金资助项目(30400399、30671917); 江苏省自然科学基金资助项目(BK2008215) |
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| 摘 要： | 为构建小鼠CD86启动子,并对其特异性调控能力进行鉴定。首先自C57BL/6J小鼠基因组中采用高保真PCR扩增小鼠CD86基因的启动子,经测序鉴定后,插入pCDNA3.1-EGFP重组质粒。然后分别转染小鼠来源细胞株:RAW264.7、NIH/3T3、P815、SP2/0、EL4细胞,观察各细胞内绿色荧光蛋白的表达。结果显示,小鼠CD86基因启动子的序列与GenBank报道一致;双酶切鉴定证实小鼠CD86启动子序列已经克隆入重组载体;小鼠CD86启动子能调控EGFP在巨噬细胞系RAW264.7细胞中表达,不能调控EGFP在NIH/3T3、P815、SP2/0和EL4非抗原提呈细胞内表达。说明成功构建小鼠CD86基因启动子,且该启动子具有特异性调控目的基因表达的活性。
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| 关 键 词： | 构建 小鼠CD86启动子 分子克隆 表达 |
Generation and characterization of the mouse CD86 promoter in vitro |
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| FAN Min-qi,GONG Wei-juan,,GONG Chun-xiang,XIAO Wei-ming,DING Yan-bing,JI Ming-chun. Generation and characterization of the mouse CD86 promoter in vitro[J]. Current Immunology, 2009, 0(5) |
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| Authors: | FAN Min-qi GONG Wei-juan GONG Chun-xiang XIAO Wei-ming DING Yan-bing JI Ming-chun |
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| Affiliation: | FAN Min-qi1,GONG Wei-juan2,3,GONG Chun-xiang2,XIAO Wei-ming3,DING Yan-bing3,JI Ming-chun2(1.Department of Reproduction,College of Animal Science and Technology,Yangzhou University,Yangzhou 225009,China,2.Department of Immunology,Medical College of Yangzhou University,Yangzhou 225001,2.Department of Gastroenterology,the Second Clinical Medical College of Yangzhou University,China) |
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| Abstract: | To generate a recombinant vector consisting of the mouse CD86 promoter and to study its expression in different tissue-derived cell lines of mice,the full-genetic sequence of mouse CD86 promoter was amplified from the genomic DNA of a C57BL/6 mouse by PCR.After identification by DNA sequencing,the PCR products were cloned into the recombined plasmid pcDNA3.1-EGFP,calling pCD86-EGFP.Then,the constructed plasmid was transfected into a variety of cell lines including RAW2674.7,NIH/3T3,P815,SP2/0,EL4 cells.The ... |
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| Keywords: | generation mouse CD86 promoter molecular cloning expression |
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