Construction of recombinant plasmid pEGFP-N1/MnSOD and its express in vitro.
- Author:
Yang YANG
1
;
Weijia KONG
Author Information
1. Department of Otolaryngology, the Union Hospital, Tongjing Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Gene Expression;
Genetic Vectors;
Plasmids;
Rats;
Rats, Sprague-Dawley;
Stria Vascularis;
cytology;
Superoxide Dismutase;
genetics;
Transfection
- From:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
2007;21(22):1034-1036
- CountryChina
- Language:Chinese
-
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.
