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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Establishing genetic transformation for comparative developmental studies in the crustacean Parhyale hawaiensis
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Establishing genetic transformation for comparative developmental studies in the crustacean Parhyale hawaiensis

机译：建立遗传转化以进行夏威夷甲壳动物的比较发育研究

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摘要

The amphipod crustacean Parhyale hawaiensis has been put forward as an attractive organism for evolutionary developmental comparisons, and considerable effort is being invested in isolating developmental genes and studying their expression patterns in this species. The scope of these studies could be significantly expanded by establishing means for genetic manipulation that would enable direct studies of gene functions to be carried out in this species. Here, we report the use of the Minos transposable element for the genetic transformation of P. hawaiensis. Transformed amphipods can be obtained from ≈30% of surviving individuals injected with both a Minos element carrying the 3xP3-DsRed fluorescent marker and with mRNA encoding the Minos transposase. Integral copies of the transposon are inserted into the host genome and are stably inherited through successive generations. We have used reporter constructs to identify a muscle-specific regulatory element from Parhyale, demonstrating that this transformation vector can be used to test the activity of cis-regulatory elements in this species. The relatively high efficiency of this transgenic methodology opens new opportunities for the direct study of as-regulatory elements and gene functions in Parhyale, allowing functional studies to be carried out beyond previously established model systems in insects.
机译：提出了两栖类甲壳类的夏威夷对虾（Parhyale hawaiensis）作为一种有吸引力的生物，用于进化发育比较，并且正投入大量精力来分离发育基因并研究它们在该物种中的表达模式。这些研究的范围可以通过建立基因操作手段来大大扩展，这将使对该物种的基因功能的直接研究成为可能。在这里，我们报告使用Minos转座元件进行夏威夷对虾的遗传转化。可以从大约30％的存活个体中获得转化的两栖动物，这些个体注射带有3xP3-DsRed荧光标记的Minos元件和编码Minos转座酶的mRNA。转座子的完整拷贝被插入宿主基因组，并通过连续的世代稳定地遗传。我们已使用报告基因构建物从Parhyale鉴定肌肉特异性调控元件，证明该转化载体可用于测试该物种中顺式调控元件的活性。这种转基因方法的相对较高的效率为直接研究Parhyale中的调节元件和基因功能开辟了新的机会，从而使功能性研究能够超越昆虫先前建立的模型系统。

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