| c-Myb截短突变体的构建及其对c-Myc蛋白表达的调节 |
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| 引用本文： | 秦玺,;程龙,;饶春明,;高凯,;杨琦,;史新昌,;徐小洁,;叶棋浓,;王军志.c-Myb截短突变体的构建及其对c-Myc蛋白表达的调节[J].生物技术通讯,2014(4):520-523. |
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| 作者姓名： | 秦玺 ;程龙 ;饶春明 ;高凯 ;杨琦 ;史新昌 ;徐小洁 ;叶棋浓 ;王军志 |
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| 作者单位： | [1]中国食品药品检定研究院生物制品检定所,北京100050; [2]军事医学科学院生物工程研究所,北京100850 |
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| 基金项目： | 国家科技重大专项（2012ZX09304010）;国家自然科学基金青年项目（31200565） |
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| 摘 要： | 目的:为了研究c-Myb蛋白各结构域的功能,构建c-Myb的6种截短突变体c-Myb-1~c-Myb-6的真核表达载体,测定不同突变体对c-Myc蛋白表达的影响。方法:以野生型c-Myb为模板,PCR扩增c-Myb-1~c-Myb-6片段,分别克隆到pcDNA3.0-FLAG载体,Western印迹检测克隆载体在293T细胞内的表达;将上述载体与野生型c-Myb转染乳腺癌细胞株MCF-7,检测其对c-Myb下游基因c-Myc表达的影响。结果:构建了c-Myb-1~c-Myb-6表达载体,与野生型c-Myb相比,c-Myb-1、c-Myb-4、c-Myb-5均能够促进MCF-7细胞中c-Myc的表达,而c-Myb-2、c-Myb-3、c-Myb-6则不能。结论:该研究为进一步探讨c-Myb在乳腺癌中的功能奠定了基础。
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| 关 键 词： | c-Myb 截短突变体 真核表达载体 c-Myc |
Construction and Regulation on c-Myc Expression of Deletion mu-tants of c-Myb |
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| Institution: | QIN Xi,CHENG Long,RAO Chun-Ming,GAO Kai,YANG Qi,SHI Xin-Chang,XU Xiao-Jie,YE Qi-Nong,WANG Jun-Zhi(1. National Institute for Food and Drug Control, Beijing 100050; 2. Beijing Institute of Biotechnology, Beijing 100850; China) |
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| Abstract: | Objective： To construct eukaryotic expression vectors containing six deletion mutants c-Myb-1~c-Myb-6 and to detect the impact of different isoforms on the the expression of c-Myc. Methods： c-Myb-1~c-Myb-6 genes were amplified from wild type of c-Myb using PCR and inserted into the vector pcNDA3.0-FLAG. West-ern blot analysis was used to detect the expression of these isoforms. Transfected these vectors and c-Myb into MCF-7 cells and detected the impact on the expression of c-Myc which regulated by c-Myb. Results： c-Myb-1~c-Myb-6 were successfully constructed. Same as wild type c-Myb, c-Myb-1, c-Myb-4 and c-Myb-5 could en-hance the expression of c-Myc in MCF-7, but c-Myb-2, c-Myb-3 and c-Myb-6 could not. Conclusion： This study could be a basis to further detect the function of c-Myb in breast cancer. |
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| Keywords: | c-Myb deletion mutants eukaryotic expression vector c-Myc |
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