| RUNX3基因真核表达载体的构建及其在乳腺癌T47D细胞中的表达 |
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| 引用本文： | 姜影,杜金荣,佟丹丹,娄阁,吴丽华,石岩.RUNX3基因真核表达载体的构建及其在乳腺癌T47D细胞中的表达[J].现代肿瘤医学,2011,19(9):1720-1723. |
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| 作者姓名： | 姜影 杜金荣 佟丹丹 娄阁 吴丽华 石岩 |
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| 作者单位： | 1. 哈尔滨医科大学附属第二医院病理科,黑龙江,哈尔滨,150086
2. 哈尔滨医科大学基础医学院病理学教研室,黑龙江,哈尔滨,150086 |
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| 基金项目： | 黑龙江省教育厅课题基金资助项目 |
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| 摘 要： | 目的:构建真核表达载体pcDNA3.1(+)-RUNX3,并在乳腺癌T47D细胞株中表达。方法:应用基因重组技术和限制性内切酶EcoRI和XhoI酶切,构建并鉴定pcDNA3.1(+)-RUNX3真核表达载体,经脂质体Lipofectamine2000介导质粒转染T47D细胞后应用逆转录-聚合酶链反应(RT-PCR)和Western blot实验,检测RUNX3在T47D细胞中的表达。结果:重组真核表达载体pcDNA3.1(+)-RUNX3经限制性内切酶EcoRI和XhoI酶切,电泳后显示1.3kb的RUNX3目的片段和5.4kb的pcDNA3.1(+)载体片段。测序证实酶切片段与gene bank中登记的RUNX3序列相同,证实pcDNA3.1(+)-RUNX3真核表达载体构建成功。经RT-PCR和Western blot检测,表明转染pcDNA3.1(+)-RUNX3的T47D细胞RUNX3阳性表达。结论:重组真核表达载体构建正确,并建立稳定表达RUNX3的T47D细胞系,从而为后续研究提供有用的细胞研究模型。
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| 关 键 词： | RUNX3基因 T47D细胞 乳腺癌 |
Construction of eukaryotic expression vector pcDNA3.1(+)-RUNX3 and its expression in human breast cancer T47D cell line |
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| JIANG Ying,DU Jinrong,TONG Dandan,LOU Ge,WU Lihua,SHI Yan.Construction of eukaryotic expression vector pcDNA3.1(+)-RUNX3 and its expression in human breast cancer T47D cell line[J].Journal of Modern Oncology,2011,19(9):1720-1723. |
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| Authors: | JIANG Ying DU Jinrong TONG Dandan LOU Ge WU Lihua SHI Yan |
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| Institution: | JIANG Ying1,DU Jinrong1,TONG Dandan2,LOU Ge1,WU Lihua1,SHI Yan11Department of Clinicalpathology,the Second Affiliated Hospital of Harbin Medical University,Harbin 150086,China,2Department of Pathology,School of Basic Medicine,Harbin Medical University,China. |
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| Abstract: | Objective: To construct the eukaryotic expression vector pcDNA3.1(+)-RUNX3 and to express it in human breast cancer T47D cell line.Methods:Eukaryotic expression vector for pcDNA3.1(+)-RUNX3 was constructed by use of recombinant DNA technique and was transfected into T47D cell by Lipofectamine 2000.The mRNA and protein expression of RUNX3 gene was detected by RT-PCR and Western blot methods.Results:The recombined eukaryotic expression vector for RUNX3 was digested with EcoRI and XhoI,and the electrophoresis ... |
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| Keywords: | RUNX3 gene T47D cell line breast cancer |
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