Anatomical Sciences Journal (Feb 2015)
Stable Transfection of pEGFP-N1-MOG Plasmid to Utilize in Multiple Sclerosis Gene Therapy
Abstract
Introduction: Multiple sclerosis (MS) is a disease of the immune system: it attacks the myelin around the axons and leaves them exposed. Destruction of myelin weakens the electrical conduction of ions and thus leads to a lack of communication in the nervous system. Methods: In the present study, we constructed recombinant plasmid and then transformed to E. coli cell. The colonies containing plasmid were selected by Colony PCR. Enzyme digestion and sequencing were utilized to approve the accuracy of the extracted plasmid of these clones. Recombinant plasmid transfect in to mesanchymal stem cells. Results: Plasmid was verified correctly. After transfection, the transcription of MOG gene and the expression of MOG protein were proved by RT-PCR, western blotting and Elisa. Conclusion: Plasmid was constructed correctly and mesenchyme stem cells were successfully transfected by transfection and protein can be expressed well, setting a proper foundation for the future studies on the transplantation of gene modified mesanchymal stem cells in order to promote Multiple sclerosis.