Lobo Neil F, Clayton John R, Fraser Malcolm J, Kafatos Fotis C, Collins Frank H
Center for Global Health and Infectious Diseases, Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46556, USA.
Nat Protoc. 2006;1(3):1312-7. doi: 10.1038/nprot.2006.221.
The ability to manipulate the mosquito genome through germ-line transformation provides us with a powerful tool for investigating gene structure and function. It is also a valuable method for the development of novel approaches to combating the spread of mosquito-vectored diseases. To date, germ-line transformation has been demonstrated in several mosquito species. Transgenes are introduced into pre-blastocyst mosquito embryos using microinjection techniques that take a few hours, and progeny are screened for the presence of a marker gene. The microinjection protocol presented here can be applied to most mosquitoes and contains several improvements over other published methods that increase the survival of injected embryos and, therefore, the number of transformants. Transgenic lines can be established in approximately 1 month using this technique.
通过种系转化来操纵蚊子基因组的能力，为我们研究基因结构和功能提供了一个强大的工具。它也是开发新型方法以对抗蚊媒疾病传播的一种有价值的方法。迄今为止，已经在几种蚊子物种中证明了种系转化。使用需要几个小时的显微注射技术将转基因导入囊胚前期蚊子胚胎，并筛选后代中标记基因的存在。这里介绍的显微注射方案可应用于大多数蚊子，并且与其他已发表的方法相比有几个改进之处，这些改进提高了注射胚胎的存活率，从而增加了转化体的数量。使用这种技术可以在大约1个月内建立转基因品系。
