Construction of recombinant plasmid pEGFP-N1/MnSOD and its express in vitro / 临床耳鼻咽喉头颈外科杂志

ABSTRACT

OBJECTIVE@#To construct eukaryotic expression plasmid of rat Mn-superoxide dismutase (MnSOD) gene pEGFP-N1/MnSOD and express it in primary culture system of marginal cells (MC) of the rats.@*METHOD@#The aimed segments were obtained from rat myocardial tissue and were inserted into a eukaryotic expression plasmid pEGFP-N1/MnSOD. The recombinant expression plasmid pEGFP-N1/MnSOD was transfected into MC by lipofectamine 2000-mediated gene transfer method, which were observed through co-Focus Fluorescence microscopy. The percentage of transfection was determined by fluorescence activated call sorting (FACS) analysis.@*RESULT@#Rat MnSOD cDNA eukaryotic expression plasmid pEGFP-N1/MnSOD have been successfully constructed. Green fluorescent protein(GFP) and MnSOD protein could be contacted in the transfected MC 48 hours after transfection. The percentage of transfection was 23.47%.@*CONCLUSION@#Rat MnSOD cDNA eukaryotic expression plasmid pEGFP-N1/MnSOD have been successfully constructed and expressed successfully in MC. The research paved the way for antioxidant gene therapy of inner ear.