Shittu Mujeeb, Steenwinkel Tessa, Koshikawa Shigeyuki, Werner Thomas
Department of Biological Sciences, Michigan Technological University, Houghton, MI 49931, USA.
Faculty of Environmental Earth Science, Hokkaido University, N10W5, Kita-ku, Sapporo, Hokkaido 060-0810, Japan.
Methods Protoc. 2020 Apr 27;3(2):31. doi: 10.3390/mps3020031.
The complex color patterns on the wings and body of () are emerging as model systems for studying evolutionary and developmental processes. Studies regarding these processes depend on overexpression and downregulation of developmental genes, which ultimately rely upon an effective transgenic system. Methods describing transgenesis in () have been reported in several studies, but they cannot be applied to due to the low egg production rate and the delicacy of the eggs In this protocol, we describe extensively a comprehensive method used for generating transgenic . Using the protocol described here, we are able to establish transgenic lines, identifiable by the expression of enhanced green fluorescent protein (EGFP) in the eye disks of larvae. The entire procedure, from injection to screening for transgenic larvae, can be completed in approximately 30 days and should be relatively easy to adapt to other non-model species, for which no white-eyed mutants exist.
（某生物）翅膀和身体上复杂的颜色图案正成为研究进化和发育过程的模型系统。关于这些过程的研究依赖于发育基因的过表达和下调，这最终依赖于一个有效的转基因系统。几项研究报告了描述（某生物）转基因的方法，但由于产卵率低和卵的脆弱性，这些方法不能应用于（另一生物）。在本方案中，我们广泛描述了一种用于产生（某生物）转基因的综合方法。使用此处描述的方案，我们能够建立转基因品系，可通过（某生物）幼虫眼盘中增强型绿色荧光蛋白（EGFP）的表达来识别。从注射到筛选转基因幼虫的整个过程大约可以在30天内完成，并且应该相对容易适用于其他没有白眼突变体的非模式（生物）物种。
