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首页> 美国卫生研究院文献>Scientific Reports >Identification of a novel strong promoter from the anhydrobiotic midge Polypedilum vanderplanki with conserved function in various insect cell lines
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Identification of a novel strong promoter from the anhydrobiotic midge Polypedilum vanderplanki with conserved function in various insect cell lines

机译：鉴定出一种新型的强启动子该新的强启动子来自水生中蚊Polypedilum vanderplanki在各种昆虫细胞系中具有保守功能

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Larvae of the African midge Polypedilum vanderplanki (Diptera: Chironomidae) show a form of extreme desiccation tolerance known as anhydrobiosis. The cell line Pv11 was recently established from the species, and these cells can also survive under desiccated conditions, and proliferate normally after rehydration. Here we report the identification of a new promoter, 121, which has strong constitutive transcriptional activity in Pv11 cells and promotes effective expression of exogenous genes. Using a luciferase reporter assay, this strong transcriptional activity was shown to be conserved in cell lines from various insect species, including S2 (Drosophila melanogaster, Diptera), SaPe-4 (Sarcophaga peregrina, Diptera), Sf9 (Spodoptera frugiperda, Lepidoptera) and Tc81 (Tribolium castaneum, Coleoptera) cells. In conjunction with an appropriate selection maker gene, the 121 promoter was able to confer zeocin resistance on SaPe-4 cells and allowed the establishment of stable SaPe-4 cell lines expressing the fluorescent protein AcGFP1; this is the first report of heterologous gene expression in this cell line. These results show the 121 promoter to be a versatile tool for exogenous gene expression in a wide range of insect cell lines, particularly useful to those from non-model insect species.
机译：非洲midPolypedilum vanderplanki（双翅目：Chironomidae）的幼虫表现出一种极端的耐旱性，称为脱水生物。最近从该物种建立了细胞系Pv11，这些细胞也可以在干燥条件下存活，并在补液后正常增殖。在这里我们报告鉴定一个新的启动子121，其在Pv11细胞中具有很强的组成型转录活性，并促进外源基因的有效表达。使用萤光素酶报告基因测定法，该强转录活性在各种昆虫物种的细胞系中均得以保留，这些昆虫物种包括S2（果蝇Drosophila melanogaster，Diptera），SaPe-4（Sarcophaga peregrina，Diptera），Sf9（Spodoptera frugiperda，Lepidoptera）和Tc81（Tribolium castaneum，鞘翅目）细胞。结合适当的选择标记基因，121启动子能够赋予SaPe-4细胞Zeocin抗性，并能建立表达荧光蛋白AcGFP1的稳定SaPe-4细胞系。这是该细胞系中异源基因表达的首次报道。这些结果表明121启动子是广泛的昆虫细胞系中外源基因表达的通用工具，对那些来自非模型昆虫物种的细胞特别有用。

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