Introduction of foreign DNA into the water flea, Daphnia magna, by electroporation
Introduction of foreign DNA into the water flea, Daphnia magna, by electroporation
复制标题
DOI：
10.1007/s10646-010-0460-9
复制
发表时间：
2010-03-01
期刊：
影响因子：
2.7
通讯作者：
Iguchi, Taisen
中科院分区：
文献类型：
--
作者：
Kato, Yasuhiko;Kobayashi, Kaoru;Iguchi, Taisen
水蚤类栖息于多种多样的水生环境中，是理解环境变化的响应和适应的良好模型，已被用作生态毒理学的标准生物之一。基因组学的最新进展改变了分析水蚤反应的工具，因为基因表达的变化可以在生物体的不动等显著不利影响出现之前观察到。因此，从基因表达水平了解生物学变化可以成为评价生物体环境响应的敏感工具之一。然而，在水蚤中没有基因操作的技术。因此，我们开发了一种基于电穿孔的基因导入技术。成功地将外源DNA导入D. magna. (1)将DNA注入血液。(2)使用非常低的电压进行电穿孔。将含有绿色荧光蛋白（GFP）的DNA注射到水蚤体内，并在活体水蚤体内检测到GFP的表达。这是首次将基因导入水蚤类动物的报道，并与新出现的基因组序列一起，将有助于扩大我们在生态毒理学中的应用。
Daphnids inhabit a diverse array of aquatic environments and they are a good model for understanding response and adaptation to environmental changes and they have been used one of standard organisms in ecotoxicology. Recent progress of genomics changed the tools for analyzing responses of daphnids, because gene expression changes can be observed before the emergence of prominent adverse effect such as immobility of the organism. Thus understanding of biological changes from gene expression level can be one of the sensitive tools for the evaluation of environmental response of organisms. However, there was no technique for genetic manipulation in daphnids. Hence, we have developed a gene introduction technique based on electroporation. There are two critical points for the successful introduction of foreign DNA into D. magna. (1) Injection of DNA into blood stream. (2) Usage of very low voltage for the electroporation. The injected DNA containing green fluorescent protein (GFP) could be introduced daphnids and the expression of GFP could be detected in living daphnids. This is the first report of gene introduction to daphnids and, together with the emerging genome sequences, will be useful for the expanding our use of daphnid in ecotoxicology.