| LHR真核表达载体构建及表达 |
| |
| 引用本文： | 林授,林德馨. LHR真核表达载体构建及表达[J]. 福建医科大学学报, 2009, 43(2): 142-144 |
| |
| 作者姓名： | 林授 林德馨 |
| |
| 作者单位： | 1. 福建省血液中心,检验科,福州,350004
2. 福建医科大学基础医学院,生物化学与分子生物学系,福州,350004 |
| |
| 基金项目： | 福建医科大学科学研究发展基金 |
| |
| 摘 要： | 目的构建黄体生成素受体（LHR）真核表达质粒载体和建立稳定高表达LHR的乳腺癌细胞株。方法KpnI／Hpa工双酶切LHR-cDNA质粒载体获得目的基因，定向克隆构建pcDNA3．1（4-）真核表达载体，酶切图谱和序列分析鉴定；重组质粒载体经脂质体转染MCF-7乳腺癌细胞，G418筛选，RT-PCR、细胞内cAMP测定，筛选鉴定高表达LHRMCF-7细胞。结果重组质粒pcDNA3．1（4-）-LHR酶切图谱分析结果、序列分析证明pcDNA3．1（4-）-LHR载体中LHR序列与GenBank的LHR基因序列完全相符。RT-PCR检测MCF-7细胞LHRmRNA，MCF-7细胞内cAMP浓度测定，证实MCF-7细胞高表达LHR。结论构建并转染pcDNA3．1（4-）-LHR真核表达载体，在MCF-7细胞中稳定表达，为探讨hCG对乳腺癌细胞的作用提供实验基础。
|
| 关 键 词： | 受体,LH 绒毛膜促性腺激素 乳腺肿瘤 克隆细胞 遗传载体 基因表达RNA,信使 |
Construction of Eukaryotic Expression Recombinant Plasmid pcDNA3.1(+)-LHR |
| |
| LIN Shou,LIN Dexin. Construction of Eukaryotic Expression Recombinant Plasmid pcDNA3.1(+)-LHR[J]. Journal of Fujian Medical University, 2009, 43(2): 142-144 |
| |
| Authors: | LIN Shou LIN Dexin |
| |
| Affiliation: | LIN Shou1,LIN Dexin2 1.Fujian Blood Center,FuZhou 350004,China,2.Department of Biochemistry and Molecular Biology,Fujian Medical University |
| |
| Abstract: | Objective To construct the eukaryotic expression recombinant plasmid pcDNA3.1(+)-LHR and to establish human breast cancer MCF-7 cell lines stably expressing exogenous LHR. Methods LHR cDNA was cloned into pcDNA3.1(+) vector,the recombinant plasmid pcDNA3.1(+)-LHR was identified by restriction enzyme digestion and DNA sequence analysis. pcDNA3.1(+)-LHR plasmid was transfected into MCF-7 cells and cell lines stably expressing exogenous LHR were established by G418 screening. The expression of LHR was detected... |
| |
| Keywords: | receptors LH chorionic gonadotropin breast neoplasms clone cells genetic vectors gene expression RNA messenger |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
