Mos1 transposon-based transformation of fish cell lines using baculoviral vectors |
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Authors: | Masako Yokoo Ryosuke Fujita Yumiko Nakajima Mamoru Yoshimizu Hisae Kasai Shin-ichiro Asano Hisanori Bando |
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Affiliation: | 1. Laboratory of Applied Molecular Entomology, Division of Applied Bioscience, Research Faculty of Agriculture, Hokkaido University, Sapporo 060-8589, Japan;2. Innate Immunity Laboratory, Graduate School of Life Science and Creative Research Institution, Hokkaido University, Sapporo 001-0021, Japan;3. Functional Genomics Group, COMB, Tropical Biosphere Research Center, University of the Ryukyus, Okinawa 903-0213, Japan;4. Faculty of Fisheries Sciences, Hokkaido University, Hakodate 041-8611, Japan |
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Abstract: | Drosophila Mos1 belongs to the mariner family of transposons, which are one of the most ubiquitous transposons among eukaryotes. We first determined nuclear transportation of the Drosophila Mos1-EGFP fusion protein in fish cell lines because it is required for a function of transposons. We next constructed recombinant baculoviral vectors harboring the Drosophila Mos1 transposon or marker genes located between Mos1 inverted repeats. The infectivity of the recombinant virus to fish cells was assessed by monitoring the expression of a fluorescent protein encoded in the viral genome. We detected transgene expression in CHSE-214, HINAE, and EPC cells, but not in GF or RTG-2 cells. In the co-infection assay of the Mos1-expressing virus and reporter gene-expressing virus, we successfully transformed CHSE-214 and HINAE cells. These results suggest that the combination of a baculovirus and Mos1 transposable element may be a tool for transgenesis in fish cells. |
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Keywords: | Baculovirus Mariner transposon Mos1 Fish cells |
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