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| 人心肌肌球蛋白轻链基因启动子驱动的GFP和Luc报告基因在人心肌细胞系中的表达 | |
| 引用本文： | 许秀芳,辛毅,汪劲松,赵莉敏,杜兰萍,葛桂玲,李娜,张颖,王世强,黄益民,罗毅.人心肌肌球蛋白轻链基因启动子驱动的GFP和Luc报告基因在人心肌细胞系中的表达[J].中国病理生理杂志,2012,28(9):1722-1728. |
| 作者姓名： | 许秀芳 辛毅 汪劲松 赵莉敏 杜兰萍 葛桂玲 李娜 张颖 王世强 黄益民 罗毅 |
| 作者单位： | 1. 首都医科大学附属北京安贞医院, 北京市心肺血管疾病研究所, 细胞与分子生物学研究室, 北京 100029; 2. 首都医科大学附属北京儿童医院心外科, 北京 100045 3. 首都医科大学附属北京安贞医院, 北京市心肺血管疾病研究所, 实验中心, 北京 100029; 4. 北京放射医学研究所, 北京 100850; 5. 北京大学生命科学学院, 北京 100871; 6. 北京中医药大学电镜室, 北京 100029 |
| 基金项目： | 北京市自然科学基金资助项目(No.5042006;No.7072012) |
| 摘 要： | 目的:构建人肌球蛋白2v基因启动子(pMLC2v)驱动的绿色荧光蛋白(GFP)和萤光素酶(Luc)报告基因慢病毒载体并观察其在人心肌细胞系(HCM)和人肺癌细胞株A549中的整合表达特征。方法:应用去毒化的人类I型免疫缺陷病毒和pMLC2v-GFP或pMLC2v-Luc报告基因构建慢病毒示踪载体,转染HCM和A549细胞,激光共聚焦显微镜及生物发光检测仪观察2种报告基因在不同细胞生长进程中的表达特征。非特异性启动子驱动的GFP(GFPC)和红色荧光蛋白(RFPC)报告基因作为对照。结果:2种细胞转染GFPC和RFPC后第3 d,都表达GFP和RFP;而HCM只在转染pMLC2v-GFP和pMLC2v-Luc 21 d后表达GFP和Luc,A549细胞不表达。结论:pMLC2v主要在培养21 d后新增殖的人心肌细胞中驱动GFP和Luc报告基因表达,这为监测干细胞向心肌细胞的分化进程提供了可靠的病理及活体示踪工具。 |
| 关 键 词： | 肌球蛋白轻链 绿色荧光蛋白 萤光素酶 心肌细胞 |
| 收稿时间： | 2012-01-16 |
Expression of GFP and Luc reporter genes driven by human myocardial myosin light chain gene promoter in human cardiomyocytes |
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| XU Xiu-fang,XIN Yi,WANG Jin-song,ZHAO Li-min,DU Lan-ping,GE Gui-ling,LI Na,ZHANG Ying,WANG Shi-qiang,HUANG Yi-min,LUO Yi.Expression of GFP and Luc reporter genes driven by human myocardial myosin light chain gene promoter in human cardiomyocytes[J].Chinese Journal of Pathophysiology,2012,28(9):1722-1728. | |
| Authors: | XU Xiu-fang XIN Yi WANG Jin-song ZHAO Li-min DU Lan-ping GE Gui-ling LI Na ZHANG Ying WANG Shi-qiang HUANG Yi-min LUO Yi |
| Institution: | 1Department of Cellular & Molecular Biology,3Experimental Center,Beijing Anzhen Hospital Affiliated to Capital Medical University,Beijing Institute of Heart,Lung & Blood Vessel Diseases,Beijing 100029,China;2Department of Cardiac Surgery,Beijing Children Hospital Affiliated to Capital Medical University,Beijing 100045,China;4Institute of Radiation Medicine,Beijing 100850,China;5School of Life Sciences,Peking University,Beijing 100871,China;6Department of Electron Microscope,Beijing University of Chinese Medicine,Beijing 100029,China.) |
| Abstract: | AIM:To construct the lentiviral vectors with green fluorescent protein(GFP) and luciferase(Luc) reporter genes driven by human myosine light chain 2v gene promoter(pMLC2v) and to investigate their expression in human cardiomyocyte(HCM) cell line and human lung cancer cell line A549. METHODS:Human pMLC2v-specific lentiviral vectors with GFP(pMLC2v-GFP) or Luc(pMLC2v-Luc) were constructed and transfected into HCM and A549 cell lines. The expression characteristics of the reporter genes were observed by confocal fluorescent microscopy and bioluminescence detection. Common(nonspecific) promoter-driven GFP(GFPC) or red fluorescent protein(RFPC) lentiviral vectors were used as controls. RESULTS:Both cell lines expressed GFP and RFP 3 days after transfected with the nonspecific vectors. HCM specifically expressed GFP and Luc 3 weeks after transfected with the pMLC2v-GFP or pMLC2v-Luc vectors. However, A549 cells didn't show the similar expression pattern. CONCLUSION:The pMLC2v-GFP and pMLC2v-Luc lentiviral vectors are specific for newly proliferative cardiomyocytes, indicating that they can be used as reliable tools for tracking the differentiation of stem cells into cardiomyocytes in vivo. |
| Keywords: | Myosin light chain Green fluorescent protein Luciferase Cardiomyocytes |
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