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A novel 2A-peptide-containing plasmid to generate stable Perkinsus marinus cells expressing organelle-targeted genes |
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| Authors: | Hirokazu Sakamoto Kiyoshi Kita Motomichi Matsuzaki |
| Institution: | 1. Department of Biomedical Chemistry, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan;2. Department of Biomedical Chemistry, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan
School of Tropical Medicine and Global Health, Nagasaki University, Nagasaki, Japan Department of Host-Defense Biochemistry, Institute of Tropical Medicine, Nagasaki University, Nagasaki, Japan |
| Abstract: | Genetic manipulation techniques for marine protists are not well-established, despite immense efforts. However, Perkinsus marinus is an exception and can be developed as a genetically tractable model organism for related protists. Here, we designed a new plasmid for P. marinus that allows two proteins from a single mRNA to be differently localized using a self-cleaving 2A peptide. This enabled us to establish a stable transfectant expressing a mitochondrially targeted fluorescent protein. The system can be applied to any protein in theory and would make a powerful tool for investigating unique organelles in P. marinus and related dinoflagellates. |
| Keywords: | alternative oxidase dinoflagellates drug selection genetic manipulation membrane-bound organelle polycistronic plasmid |