首页|期刊导航|南京医科大学学报（英文版）|Constructing retroviral vector carrying green fluorescent protein (GFP) and investigating the expression of GFP in primary rat myoblast
南京医科大学学报（英文版）2006，Vol.20Issue(4)：197-200,4.
Constructing retroviral vector carrying green fluorescent protein (GFP) and investigating the expression of GFP in primary rat myoblast
Constructing retroviral vector carrying green fluorescent protein (GFP) and investigating the expression of GFP in primary rat myoblast
摘要
Abstract
Objective: To construct green fluorescent protein (GFP) retroviral vector (pLgXSN), and to investigate the expression of GFP in primary rat myoblast. Methods: GFP cDNA was subcloned into the plasmid pLgXSN, and the recombinant vector was transfected into packaging cell PT67. G418 was used to select positive colony. Myoblasts were infected by a high-titer viral supernatant. The recombinant retroviral plasmid vector was identified by restriction endonuclease analysis and DNA sequence analysis. Confocal microscopy and flow cytometry were used to detect the expression of GFP. Results: The GFP cDNA sequence was identical to that of GenBank. Recombinant retroviral plasmid vector pLgGFPSN was constructed successfully. The titer of the packaged recombinant retrovirus was 1 × 106 cfu/ml. Bright green fluorescence of the transfected cells was observed under confocal microscope 48 h after transfection. The transfection rate was 33%. The effective expression of GFP in myoblast infected by recombinant retrovirus lasted for 6 weeks. Conclusion: GFP gene could be effectively and stably expressed in myoblast, which suggests that GFP could act as a marker for studies on myoblast.关键词
green fluorescent protein/transfection/myoblastKey words
green fluorescent protein/transfection/myoblast分类
医药卫生引用本文复制引用
Shuling Rong,Yongxin Lu,Yuhua Liao,Xiaolin Wang,Xiaoqing Li,Jiahua Zhang ,Yanli He..Constructing retroviral vector carrying green fluorescent protein (GFP) and investigating the expression of GFP in primary rat myoblast[J].南京医科大学学报（英文版）,2006,20(4):197-200,4.基金项目
National 863 Program of China(2002AA745070) （2002AA745070）
