| 小鼠4-1BBL基因真核表达载体构建及其在前列腺癌细胞中的表达 |
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| 引用本文： | 冯正秋,刘修恒,江波涛,翁小东,匡幼林.小鼠4-1BBL基因真核表达载体构建及其在前列腺癌细胞中的表达[J].武汉大学学报(医学版),2009,30(5). |
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| 作者姓名： | 冯正秋 刘修恒 江波涛 翁小东 匡幼林 |
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| 作者单位： | 武汉大学人民医院泌尿外科,湖北,武汉,430060 |
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| 基金项目： | 国家自然科学基金资助项目(编号:30672107) |
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| 摘 要： | 目的:构建含小鼠4-1BBL编码区cDNA序列的真核表达载体,并检测其在前列腺癌细胞株(PC-3)中的表达。方法:将目的基因m4-1BBL全长cDNA克隆到真核表达载体pcDNA3.1(+),获得m4-1BBL定向插入重组子pcDNA3.1(+)m4-1BBL,采用限制性内切酶和测序鉴定是否成功构建。用脂质体法将重组质粒转入PC-3细胞,逆转录-聚合酶链反应(RT-PCR)和Western blot检测m4-1BBL在PC-3中的表达。结果:酶切鉴定和序列分析证实,重组质粒含有小鼠4-1BBL全长cDNA编码序列,转染实验表明m4-1BBL基因能在PC-3细胞中表达。结论:小鼠4-1BBL全长cDNA基因真核表达载体构建及其在PC-3细胞中的表达均获成功,为进一步研究莫定了基础。
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| 关 键 词： | 小鼠4-1BBL 基因克隆 真核表达 共刺激分子 |
Construction of Eukaryotic Expression Vectors Containing Mouse 4-1BBL Gene and Its Expression in Prostate Cancer Cells |
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| FENG Zhengqiu,LIU Xiuheng,JIANG Botao,WENG Xiaodong,KUANG Youlin.Construction of Eukaryotic Expression Vectors Containing Mouse 4-1BBL Gene and Its Expression in Prostate Cancer Cells[J].Medical Journal of Wuhan University,2009,30(5). |
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| Authors: | FENG Zhengqiu LIU Xiuheng JIANG Botao WENG Xiaodong KUANG Youlin |
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| Institution: | FENG Zhengqiu,LIU Xiuheng,JIANG Botao,WENG Xiaodong,KUANG YoulinDept.of Urology,Renmin Hospital of Wuhan University,Wuhan 430060,China |
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| Abstract: | Objective: To construct a eukaryotic expression vector containing the coding region of mouse 4-1BBL cDNA and detect its expression in prostate cancer cell strain(PC-3).Methods: The murine full-length 4-1BBL cDNA was obtained and cloned into the pcDNA3.1(+) eukaryotic expression vector.The recombinant pcDNA3.1(+)-m4-1BBL was identified by restriction enzyme digestion and sequencing.Then the recombinated plasmid pcDNA3.1(+)-m4-1BBL was transfected into PC-3 cells using Lipofectamine 2000.The m4-1BBL transfete... |
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| Keywords: | Mouse 4-1BBL Gene Cloning Eukaryotic Expression Co-Stimulating Factor |
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