Abstract：
Transgenesis in Drosophila is an essential approach to studying gene function at the organism level. Embryo microinjection is a crucial step for the construction of transgenic flies. Microinjection requires some types of equipment, including a microinjector, a micromanipulator, an inverted microscope, and a stereo microscope. Plasmids isolated with a plasmid miniprep kit are qualified for microinjection. Embryos at the pre-blastoderm or syncytial blastoderm stage, where nuclei share a common cytoplasm, are subjected to microinjection. A cell strainer eases the process of dechorionating embryos. The optimal time for dechorionation and desiccation of embryos needs to be determined experimentally. To increase the efficiency of embryo microinjection, needles prepared by a puller need to be beveled by a needle grinder. In the process of grinding needles, we utilize a foot air pump with a pressure gauge to avoid the capillary effect of the needle tip. We routinely inject 120-140 embryos for each plasmid and obtain at least one transgenic line for around 85% of plasmids. This article takes the phiC31 integrase-mediated transgenesis in Drosophila as an example and presents a detailed protocol for embryo microinjection for transgenesis in Drosophila.
摘要：
果蝇的转基因是在生物体水平上研究基因功能的重要方法。胚胎显微注射是构建转基因果蝇的关键步骤。显微注射需要一些类型的设备，包括一个微型注射器，一个微操纵器,倒置显微镜，和一个立体显微镜。用质粒小量制备试剂盒分离的质粒有资格进行显微注射。前胚盘或合胞胚盘阶段的胚胎，细胞核共享一个共同的细胞质，进行显微注射。细胞过滤器简化了胚胎的去亲过程。胚胎脱泡和干燥的最佳时间需要通过实验确定。为了提高胚胎显微注射的效率，拉拔器准备的针需要用针磨机斜切。在磨针的过程中，我们使用带有压力表的脚踏气泵，以避免针尖的毛细作用。我们通常为每个质粒注射120-140个胚胎，并获得至少一个约85％质粒的转基因系。本文以果蝇中phiC31整合酶介导的转基因为例，提出了果蝇中胚胎显微注射转基因的详细方案。
