Transgenesis in Drosophila melanogaster.
Transgenesis in Drosophila melanogaster.
复制标题
DOI：
10.1007/978-1-60327-019-9_1
复制
发表时间：
2009-01-01
期刊：
影响因子：
--
通讯作者：
Ringrose, Leonie
中科院分区：
文献类型：
--
作者：
Ringrose, Leonie
果蝇的转基因依赖于胚胎的直接显微注射和随后存活的成虫的杂交。对单个果蝇进行杂交以筛选转基因事件的必要性将有用的转基因技术的范围限制在那些整合频率非常高的转基因技术上，因此大约每10只存活的成年果蝇中就有1只携带转基因。直到最近，只有随机的P-元件转基因符合这些标准。然而，最近的进展使同源重组和定点整合达到并超过了这种效率水平。在果蝇的所有转基因技术中，胚胎显微注射是核心步骤。本章给出了显微注射的详细方案，旨在使读者能够将其用于定点整合和P-元素转基因。
Transgenesis in Drosophila melanogaster relies upon direct microinjection of embryos and subsequent crossing of surviving adults. The necessity of crossing single flies to screen for transgenic events limits the range of useful transgenesis techniques to those that have a very high frequency of integration, so that about 1 in 10 to 1 in 100 surviving adult flies carry a transgene. Until recently, only random P-element transgenesis fulfilled these criteria. However, recent advances have brought homologous recombination and site-directed integration up to and beyond this level of efficiency. For all transgenesis techniques in Drosophila melanogaster, microinjection of embryos is the central procedure. This chapter gives a detailed protocol for microinjection, and aims to enable the reader to use it for both site-directed integration and for P-element transgenesis.