Laboratory of Applied Molecular Entomology, Division of Applied Bioscience, Research Faculty of Agriculture, Hokkaido University, Sapporo 060-8589, Japan.
Biochem Biophys Res Commun. 2013 Sep 13;439(1):18-22. doi: 10.1016/j.bbrc.2013.08.037. Epub 2013 Aug 17.
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.
果蝇 Mos1 属于转座子的 mariner 家族，它是真核生物中最普遍存在的转座子之一。我们首先确定了果蝇 Mos1-EGFP 融合蛋白在鱼类细胞系中的核运输，因为这是转座子功能所必需的。接下来，我们构建了携带果蝇 Mos1 转座子或位于 Mos1 反向重复之间的标记基因的重组杆状病毒载体。通过监测病毒基因组中编码的荧光蛋白的表达来评估重组病毒对鱼类细胞的感染性。我们在 CHSE-214、HINAE 和 EPC 细胞中检测到转基因表达，但在 GF 或 RTG-2 细胞中未检测到。在表达 Mos1 的病毒和报告基因表达病毒的共感染实验中，我们成功地转化了 CHSE-214 和 HINAE 细胞。这些结果表明，杆状病毒和 Mos1 转座元件的组合可能是鱼类细胞转基因的一种工具。
