Mos1 transposon-based transformation of fish cell lines using baculoviral vectors
Mos1 transposon-based transformation of fish cell lines using baculoviral vectors
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使用杆状病毒载体对鱼细胞系进行基于 Mos1 转座子的转化
DOI：
10.1016/j.bbrc.2013.08.037
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发表时间：
2013
影响因子：
3.1
通讯作者：
Hironori Bando
中科院分区：
文献类型：
--
作者：
Masako Yokoo、Ryosuke Fujita、Yumiko Nakajima、Mamoru Yoshimizu、Hisae Kasai、Shin-ichiro Asano;Hironori Bando
果蝇Mos1属于水手类转座子家族，是真核生物中最普遍的转座子之一。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，因为它是转座子功能所必需的。接下来，我们构建了含有果蝇Mos1转座子或位于Mos1反向重复序列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。我们检测到转基因在CHSE-214、HINAE和EPC细胞中的表达，但在GF和RTG-2细胞中没有表达。在表达Mos1的病毒和表达报告基因的病毒的混合感染实验中，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转座子的结合可能是鱼类细胞转基因的工具。
Drosophila Mos1belongs to the mariner family of transposons, which are one of the most ubiquitous transposons among eukaryotes. We first determined nuclear transportation of theDrosophila Mos1-EGFP fusion protein in fish cell lines because it is required for a function of transposons. We next constructed recombinant baculoviral vectors harboring theDrosophila Mos1transposon or marker genes located betweenMos1inverted 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 theMos1-expressing virus and reporter gene-expressing virus, we successfully transformed CHSE-214 and HINAE cells. These results suggest that the combination of a baculovirus andMos1transposable element may be a tool for transgenesis in fish cells.