Ittner Lars M, Götz Jürgen
Alzheimer's & Parkinson's Disease Laboratory, Brain & Mind Research Institute, University of Sydney, 100, Mallett Street, Camperdown, New South Wales 2050, Australia.
Nat Protoc. 2007;2(5):1206-15. doi: 10.1038/nprot.2007.145.
Transgenic mice have been instrumental in dissecting the role of various neuronal proteins under both physiological and pathological conditions. Pronuclear injection is the most widely used protocol for the generation of transgenic mice. Here, we describe all steps involved from DNA purification to the set up of a mouse colony including vasectomy, injection of the DNA into a donor zygote, transfer of injected zygotes into recipient foster mice, screening of offspring and establishment of transgenic mouse lines. We discuss the use of neuron-specific promoters to express proteins with a role in Alzheimer disease. Transgenic expression of a truncated form of the microtubule-associated protein tau (delta tau) is used as an example for the anticipated results.
转基因小鼠在剖析各种神经元蛋白在生理和病理条件下的作用方面发挥了重要作用。原核注射是生成转基因小鼠最广泛使用的方案。在此，我们描述了从DNA纯化到建立小鼠群体所涉及的所有步骤，包括输精管切除术、将DNA注射到供体受精卵中、将注射后的受精卵转移到受体寄养小鼠中、筛选后代以及建立转基因小鼠品系。我们讨论了使用神经元特异性启动子来表达在阿尔茨海默病中起作用的蛋白质。以微管相关蛋白tau的截短形式（δtau）的转基因表达为例来说明预期结果。
