Diegelmann Soeren, Bate Michael, Landgraf Matthias
Department of Zoology; University of Cambridge; Cambridge, United Kingdom.
Fly (Austin). 2008 Jul-Aug;2(4):236-9. doi: 10.4161/fly.6817.
Studies on the formation of connections in the developing nervous system are greatly aided by methods that permit the differential visualisation and manipulation of pre- and postsynaptic partner neurons. This has been facilitated by the advent of the LexA-based, GAL4/UAS-independent, binary expression system. On the molecular side, the introduction of DNA sequences into expression vectors has been simplified by the Invitrogen Gateway cloning technology. We have developed cloning vectors that combine the Gateway cloning technology with the LexA-based genetic expression system. These vectors facilitate the creation of driver and reporter constructs for the generation of Drosophila transgenic lines for the new LexA-based binary transcriptional system. We further report a new LexA::GAD sensory neuron driver and a red fluorescent membrane targeted lexAop reporter designed to complement the existing GFP-based lexAop reporter. Using these transgenic lines we have been able to differentially label motor and sensory neuron projections in the ventral nerve cord of Drosophila larvae.
在发育中的神经系统中，对突触前和突触后伙伴神经元进行差异可视化和操作的方法极大地促进了对连接形成的研究。基于LexA的、独立于GAL4/UAS的二元表达系统的出现推动了这一进展。在分子层面，Invitrogen Gateway克隆技术简化了将DNA序列引入表达载体的过程。我们开发了将Gateway克隆技术与基于LexA的基因表达系统相结合的克隆载体。这些载体有助于构建驱动和报告构建体，用于为基于LexA的新二元转录系统生成果蝇转基因品系。我们还报告了一种新的LexA::GAD感觉神经元驱动体和一种红色荧光膜靶向lexAop报告基因，旨在补充现有的基于GFP的lexAop报告基因。利用这些转基因品系，我们能够对果蝇幼虫腹神经索中的运动和感觉神经元投射进行差异标记。
