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| 重组质粒pENTR-CMV-EGFP-hsa-mir-16-1/15a的构建与表达 | |
| 引用本文： | 方潇碧,张春鸿,黄亚,林森,黄振校,吴丽萍,施清圆,李文峰,廖志苏.重组质粒pENTR-CMV-EGFP-hsa-mir-16-1/15a的构建与表达[J].实用医学杂志,2011,27(12). |
| 作者姓名： | 方潇碧 张春鸿 黄亚 林森 黄振校 吴丽萍 施清圆 李文峰 廖志苏 |
| 作者单位： | 1. 温州医学院附属第一医院耳鼻咽喉科,325000 2. 浙江省温州市第八人民医院耳鼻咽喉科,325000 3. 温州医学院附属第一医院放疗科,325000 |
| 基金项目： | 浙江省科技计划项目基金资助项目，浙江省温州市科技计划项目基金资助项目，浙江省温州市鹿城区科技计划项目基金资助项目 |
| 摘 要： | 目的:构建针对人鼻咽癌CNE-2Z细胞Bcl-2基因pENTR-CMV-EGFP-hsa-mir-16.1/15a真核表达质粒,转染至CNE-2Z细胞并检测其表达.方法:采用PCR法从重组质粒PGH-16-1/15a中获得16-1/15a-X2370G全长序列,在T4 DNA Ligase连接酶作用下连接入重组载体pENTR-CMV-EGFP.重组质粒经酶切及测序鉴定.将构建成功的重组质粒转染入人鼻咽癌细胞株CNE-2Z,用荧光显微镜观察转染结果.结果:重组质粒pENTR-CMV-EGFP-hsa-mir-16-1/15a经酶切与测序证实构建成功,转染至鼻咽癌CNE-2Z细胞后,荧光显微镜观察证实该重组质粒能在CNE-2Z中表达.结论:成功构建真核表达质粒DENTR-CMV-EGFP-hsa-mir-16-1/15a,并在鼻咽癌CNE-2Z细胞中得到表达,可用于进一步检测其抗肿瘤机制. |
| 关 键 词： | 质粒 构建 表达 |
Construction and expression of recombinant plasmid pENTR-CMV-EGFP -hsa-mir-16-1/15a |
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| FANG Xiao-bi,ZHANG Chun-hong,HUANG Ya,LIN Sen,HUANG Zhen-xiao,WU Li-ping,SHI Qing-yuan,LI Wen-feng,LIA Zhi-su.Construction and expression of recombinant plasmid pENTR-CMV-EGFP -hsa-mir-16-1/15a[J].The Journal of Practical Medicine,2011,27(12). | |
| Authors: | FANG Xiao-bi ZHANG Chun-hong HUANG Ya LIN Sen HUANG Zhen-xiao WU Li-ping SHI Qing-yuan LI Wen-feng LIA Zhi-su |
| Institution: | FANG Xiao-bi*,ZHANG Chun-hong,HUANG Ya,LIN Sen,HUANG Zhen-xiao,WU Li-ping,SHI Qing-yuan,LI Wen-feng,LIAO Zhi-su.*Department of Otolaryngology,The First Affiliated Hospital of Wenzhou Medical College,Wenzhou 325000,China |
| Abstract: | Objective To construct the eukaryotic expression plasmid pENTR-CMV-EGFP-hsa-mir-16-1/15a of human Bcl-2 gene,transfect it into nasopharyngeal carcinoma cells CNE-2Z and detect its expression.Methods The full-length cDNA of 16-1/15a-X2370G gene was amplified by PCR from the recombinant plasmid PGH-16-1/15a,then was linked with the recombinant vector pENTR-CMV-EGFP by T4 DNA Ligase.The recombinant plasmid was identified by restriction enzyme digestion and sequence analysis.Then the plasmid was correctly trans... |
| Keywords: | 16-1/15a-X2370G CNE-2Z |
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