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首页> 外文会议>12th Symposium on Bioluminescence and Chemiluminescence, Apr 5-9, 2002, Robinson College, University of Cambridge, UK >INTRODUCTION OF GFP-CONSTRUCT INTO MEDAKAFISH USING THE PARTICLE GUN METHOD
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INTRODUCTION OF GFP-CONSTRUCT INTO MEDAKAFISH USING THE PARTICLE GUN METHOD

机译：粒子枪法将GFP构建体引入介意

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摘要

Medakafish (Oryzias latipes) belongs to a group of small oviparous teleost fishes that inhabit fresh water in Asian countries including Japan. Medakafish has been widely used to assess the biological effects of environmental chemicals, due to its high sensitivity and ease of breeding. Also the transparency of the medakafish body enables us to investigate the expression of genes tagged by appropriate markers through its entire development, from very early in embryogenesis to a mature adult. Experimental procedures to introduce foreign genes into medakafish have been reported using electroporation and/or microinjection. Microinjection is widely used in gene transfer experiments. However, it requires high technical sophistication, and is not readily applicable to the routine introduction of several gene constructs. Electroporation can be easily applied to gene transfer experiments, but with a low efficiency. Therefore, we attempted to develop an experimental procedure to introduce a foreign gene without high technical sophistication but at reasonably high efficiency. A possible technique for this purpose was the particle gun method. However, the particle gun method has not yet been successfully applied to medakafish. One possible cause of the previous failures may have been the inappropriate combination of critical parameters required for the successful introduction of foreign genes. In this report, we describe technical data which should be useful for determining such a combination of critical parameters to introduce a foreign gene into the fertilized egg of medakafish using the particle gun method.
机译：Medakafish（Oryzias latipes）属于一组小型卵生硬骨鱼类，它们生活在包括日本在内的亚洲国家的淡水中。 Medakafish由于其高灵敏度和易于繁殖而被广泛用于评估环境化学品的生物效应。同样，medakafish体的透明性使我们能够研究整个胚胎发育过程中从适当的标记物标记的基因的表达，从胚胎发生的早期到成年。已经报道了使用电穿孔和/或显微注射将外来基因引入到aka鱼中的实验程序。显微注射广泛用于基因转移实验。然而，它需要高度的技术复杂性，并且不容易应用于几种基因构建体的常规导入。电穿孔可以很容易地应用于基因转移实验，但是效率很低。因此，我们试图开发一种实验方法来引入外源基因，而没有高技术复杂度，但是以相当高的效率。为此目的可能的技术是粒子枪法。然而，粒子枪方法尚未成功地应用于蓝aka鱼。先前失败的一个可能原因可能是成功引入外源基因所需的关键参数的不适当组合。在本报告中，我们描述了一些技术数据，这些技术数据对于确定使用粒子枪法将外来基因引入medakafish受精卵的关键参数的组合应该是有用的。

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