| چکیده فارسی مقاله |
Results This study shows the prepared gene construct is inducible by glucose. Gene expression of preproinsulin was observed in muscle tissue of rabbits. Conclusions These finding indicated that research in diabetes mellitus gene therapy could be performed on larger animals. Objectives The purpose of this study was to construct glucose inducible insulin gene plasmid and use it in the muscle tissue of the rabbit. Materials and Methods To achieve this goal, the preproinsulin, metallothionein2A promoter and the response element to carbohydrate genes were cloned into pBIND plasmid by standard cloning methods, to construct pBINDMTChIns. The gene cloning products were confirmed by the polymerase chain reaction (PCR) and restriction enzyme digestion template. The recombinant plasmid, containing the preproinsulin gene, was transferred into NIH3T3 cells and insulin gene expression was evaluated by reverse transcriptase PCR and western blotting techniques. Plasmid naked DNA containing the preproinsulin gene was injected into the rabbits’ thigh muscles, and its expression was confirmed by western blotting method. Background Diabetes mellitus type 1, formerly called insulin-dependent diabetes, is one of the autoimmune diseases where insulin-producing cells are destroyed by autoimmune response via T cells. The new approaches in treatment of diabetes are using the stem cells, cell transplantation of islet β cell, gene transfer by virus based plasmids, and non-viral gene constructs. |
| چکیده انگلیسی مقاله |
Results This study shows the prepared gene construct is inducible by glucose. Gene expression of preproinsulin was observed in muscle tissue of rabbits. Conclusions These finding indicated that research in diabetes mellitus gene therapy could be performed on larger animals. Objectives The purpose of this study was to construct glucose inducible insulin gene plasmid and use it in the muscle tissue of the rabbit. Materials and Methods To achieve this goal, the preproinsulin, metallothionein2A promoter and the response element to carbohydrate genes were cloned into pBIND plasmid by standard cloning methods, to construct pBINDMTChIns. The gene cloning products were confirmed by the polymerase chain reaction (PCR) and restriction enzyme digestion template. The recombinant plasmid, containing the preproinsulin gene, was transferred into NIH3T3 cells and insulin gene expression was evaluated by reverse transcriptase PCR and western blotting techniques. Plasmid naked DNA containing the preproinsulin gene was injected into the rabbits’ thigh muscles, and its expression was confirmed by western blotting method. Background Diabetes mellitus type 1, formerly called insulin-dependent diabetes, is one of the autoimmune diseases where insulin-producing cells are destroyed by autoimmune response via T cells. The new approaches in treatment of diabetes are using the stem cells, cell transplantation of islet β cell, gene transfer by virus based plasmids, and non-viral gene constructs. |
| نویسندگان مقاله |
hossein piri - cellular and molecular research center, qazvin university of medical sciences, qazvin, ir iran; department of biochemistry and genetics, school of medicine, qazvin university of medical sciences, qazvin, ir iran
bahram kazemi - cellular and molecular biology research center, shahid beheshti university of medical sciences, tehran, ir iran; department of biotechnology, school of medicine, shahid beheshti university of medical sciences, tehran, ir iran
iraj khodadadi - department of biochemistry and nutrition, school of medicine, hamadan university of medical sciences, hamadan, ir iran
maryam javadi - department of nutrition and dietary therapy, school of medicine, qazvin university of medical sciences, qazvin, ir iran
mojgan bandehpour - cellular and molecular biology research center, shahid beheshti university of medical sciences, tehran, ir iran; department of biotechnology, school of medicine, shahid beheshti university of medical sciences, tehran, ir iran
jamshid karimi - department of biochemistry and nutrition, school of medicine, hamadan university of medical sciences, hamadan, ir iran
amir ziaee - department of endocrinology, qazvin university of medical sciences, qazvin, ir iran; qazvin metabolic disease research center, qazvin university of medical sciences, qazvin, ir iran
amaneh koochaki - cellular and molecular biology research center, shahid beheshti university of medical sciences, tehran, ir iran; department of biotechnology, school of medicine, shahid beheshti university of medical sciences, tehran, ir iran
ali torabi - influenza unit, pasteur institute of iran, tehran, ir iran
mohammad taghi goodarzi - research center for molecular medicine, hamadan university of medical sciences, hamadan, ir iran; research center for molecular medicine, hamadan university of medical sciences, hamadan, ir iran. tel 98-8138380462, fax 98-8138380208
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