| 人miR-106b的克隆及其慢病毒表达载体构建 |
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| 引用本文： | 郑林,王爽,丁彦青.人miR-106b的克隆及其慢病毒表达载体构建[J].中国热带医学,2011,11(6):651-653. |
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| 作者姓名： | 郑林 王爽 丁彦青 |
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| 作者单位： | 南方医科大学南方医院病理科,广东,广州,510515 |
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| 基金项目： | 国家自然科学基金(81071735);国家自然科学青年基金(81000953); 广东省高校科技创新重点项目(cxzd1016); 广东省自然科学基金博士启动项目(10451051501004710) |
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| 摘 要： | 目的克隆微小RNAhsa—miR-106b并构建其慢病毒表达载体。方法将PCR扩增得到的miR-106b前体序列和pLVTHM载体经双酶切后连接，产生pLVTHM—miR-106b慢病毒表达载体，双酶切后测序鉴定，筛选阳性克隆。用pLVTHM—miR-106b、psPAX2和pMD2．G质粒共转染包装细胞293FT，包装产生慢病毒。结果经双酶切鉴定和测序证实，成功构了miR-106b的慢病毒表达载体pLVTHM—miR-106b。倒置荧光显微镜下观察可见包装细胞293FT呈绿色荧光。结论成功构建了has—miR-106b的慢病毒表达载体，为深入研究miR-106b的生物学功能奠定基础。
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| 关 键 词： | hsa—miR-106b 肿瘤 慢病毒 |
Cloning of hsa-miR-106b and construction of its lentiviral expression vector |
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| ZHENG Lin,WANG Shuang,DING Yan-qing.Cloning of hsa-miR-106b and construction of its lentiviral expression vector[J].China Tropical Medicine,2011,11(6):651-653. |
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| Authors: | ZHENG Lin WANG Shuang DING Yan-qing |
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| Institution: | ZHENG Lin,WANG Shuang,DING Yan-qing.(Department of Pathology,Nanfang Hospital,Southern Medical University,GuangZhou 510515,guandong,P.R.China) |
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| Abstract: | Aim To construct a lentiviral expression vector for hsa-miR-106b.Methods The pre-mir-106b amplified by PCR was inserted into pLVTHM.The recombinant plasmid pLVTHM-miR-106b was confirmed by restriction endonuclease analysis and DNA sequence.293FT cells were cotransfected with lentivirus vector pLVTHM-miR-106b,psPAX2 and pMD2.G.All virus stocks were produced by calcium phosphate-mediated transfection.Results Restriction enzyme digestion and DNA sequencing demonstrated that the lentivirus vector pLVTHM-miR-106... |
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| Keywords: | Hsa-miR-106b Tumor Lentivirus |
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