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| 真核表达载体pcDNA3.1(-)+KG的构建及在HeLa细胞中的表达 | |
| 引用本文： | 王洪振,王晓光,翟雷. 真核表达载体pcDNA3.1(-)+KG的构建及在HeLa细胞中的表达[J]. 生物技术通报, 2010, 0(1) |
| 作者姓名： | 王洪振 王晓光 翟雷 |
| 作者单位： | 1. 吉林师范大学生命科学学院,四平,136000 2. 长春师范学院生命科学学院,长春,130032 3. 东北师范大学遗传与细胞研究所,长春,130024 |
| 基金项目： | 国家自然科学基金，吉林师范大学博士科研启动项目 |
| 摘 要： | 为了将绿色荧光蛋白(green fluorescent protein,GFP)引入细胞核内,采用两轮PCR方法从原先克隆在pcD-NA3.1(-)+GFP载体中将GFP编码序列扩增出来并引入Kozak序列和核定位信号,使用常规酶切和连接方法将其重组至pUCm-T克隆载体中,再将目的片段重组至pcDNA3.1(-)中,对阳性克隆进行酶切、PCR和测序鉴定后,构建了带有Kozak序列和核定位信号的绿色荧光蛋白(GFP)真核表达载体pcDNA3.1(-)+KG。真核表达载体pcDNA3.1(-)+KG被转染试剂Su-perfect转染至HeLa细胞中,绿色荧光蛋白基因在HeLa细胞中得到表达而且在细胞核中观察到绿色荧光。该研究以绿色荧光蛋白为标记初步建立了活体观察真核细胞核动态变化的研究体系。 |
| 关 键 词： | Kozak序列 核定位信号 绿色荧光蛋白 真核表达载体 |
Construction of Eukaryotic Expression Vector pcDNA3.1(-)+KG Containing Green Fluorescent Protein and Its Expression in HeLa Cell |
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| Wang Hongzhen,Wang Xiaoguang,Zhai Lei. Construction of Eukaryotic Expression Vector pcDNA3.1(-)+KG Containing Green Fluorescent Protein and Its Expression in HeLa Cell[J]. Biotechnology Bulletin, 2010, 0(1) | |
| Authors: | Wang Hongzhen Wang Xiaoguang Zhai Lei |
| Affiliation: | Wang Hongzhen1 Wang Xiaoguang2 Zhai Lei3(1College of Life Science,Jilin Normal University,Siping 136000,2 College of Life Science,Changchun NormalUniversity,Changchun 130032,3Institute of Cytology , Genetics,Northeast Normal University,Changchun 130024) |
| Abstract: | To introduce green fluorescent protein into nucleus,an eukaryotic expression vector pcDNA3.1(-)+KG containing green fluorescent protein gene with kozak sequence and NLS(nuclear localization signal)was constructed.GFP sequence was amplified by two rounds of PCR from pcDNA3.1(-)+GFP vector with simultaneously being attached Kozak sequence and NLS,and then cloned into pUCm-T vector following the routine procedures.The interest gene fragment was recombinated into eukaryotic expression vector pcDNA3.1(-)to const... |
| Keywords: | Kozak sequence Nuclear localization signal Green fluorescent protein Eukaryotic expression vector |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |