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| Construction and Functional Identification of Recombinant Adeno-Associated Virus Type 9 Containing Human Carabin Gene |
| ZHOU Dianru, CHENG Kuoju, et al |
| Dazhou City Hospital of Integrated Traditionnal and Western Medicine, Sichuan Dazhou 635000, China |
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Abstract Objective: To construct recombinant adeno-associated virus type 9 containing human carabin gene and identify its function. Methods: PUC57-carabin and pAAV9-IRES-ZsGreen plasmids was digested by EcoRI + XhoI,then the enzymatic digestion product was ligated by T4DNA Ligase. The pAAV9-carabin-IRES-ZsGreen plasmid was screened in JM109 cells. The pAAV9-carabin-IRES-ZsGreen overexpression vector plasmid was transfected into 293AAV cells using high-efficiency transfection HET kit reagent to package rAAV9-carabin-IRES-ZsGreen. The viral titer was detected by Q-PCR. AC16 cells were infected with rAAV9-carabin-IRES-ZsGreen p,then the carabin protein expression was detected by western blot. Results: The pAAV9-carabin-IRES-ZsGreen overexpression plasmid was successfully constructed. RAAV9-carabin-IRES- ZsGreen was successfully packaged with a titer of 5.09E+12 vg/ml. The expression of carabin protein was significantly increased after AC16 cells were infected. Conclusion: RAAV9-carabin-IRES-ZsGreen has been successfully constructed and can express carabin efficiently in cardiomyocytes.
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2021, Vol. 27 
