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- W2188517640 abstract "Objective: To construct recombinant adeno-associated virus carrying human α-synuclein gene under the control of cytomegalovirus (CMV) or murine myelin basic protein (MBP) promoter and to measure the virus titer and verify the recombination. Methods: Human α-Synuclein and MBP cDNA were amplified by polymerase chain reaction (PCR), digested and subcloned into shuttle plasmid pSNAV2.0 to obtain recombinant plasmid pSNAV2-CMV-α-Synuclein and pSNAV2-MBP-α-Synuclein. The resulting recombinant plasmids were co-transfected into the 293T cells by calcium-phosphate precipitation method to complete rAAV2-α-Synuclein packaging. The titer of the recombinant rAAV2-α-Synuclein was determined by dot-blot assay. The recombinant rAAV-α-Synuclein was verified by PCR of the exogenous interest genes. Results: The recombinant AAV vector containing α-Synuclein genes under the control of CMV or MBP promoter were successfully constructed. The physical particle titer of rAAV2-CMV-α-Synuclein and rAAV2-MBP-α-Synuclein were respectively 5.6×10 11 vg/ml and 3.1×10 11 vg/ml. The recombinant virus was confirmed by PCR of exogenous α-Synuclein and MBP promoter gene. Conclusion: rAAV2-α-Synuclein was successfully constructed with a high virus titer, which may offer foundation for in vitro and in vivo experiments and provide a new method for the study of pathogenetic mechanisms and exploration of new therapeutic targets of particular relevance to human neurodegenerative disorders. (Tian Huijun, Wang Xiangqing, Lang Senyang. Construction of a recombinant AAV vector encoding human alpha-synuclein gene with myelin basic protein Promoter. Life Sci J 2014;11(3):214-219). (ISSN:1097-8135). http://www.lifesciencesite.com . 30" @default.
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- W2188517640 date "2014-01-01" @default.
- W2188517640 modified "2023-09-26" @default.
- W2188517640 title "Construction of a recombinant AAV vector encoding human alpha-synuclein gene with myelin basic protein Promoter" @default.
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