Gharbiah Maey, Cooley James, Leise Esther M, Nakamoto Ayaki, Rabinowitz Jeremy S, Lambert J David, Nagy Lisa M
Department of Molecular and Cellular Biology, University of Arizona, Tucson, AZ 85721, USA.
Cold Spring Harb Protoc. 2009 Apr;2009(4):pdb.prot5187. doi: 10.1101/pdb.prot5187.
The marine gastropod Ilyanassa obsoleta is a long-standing and very useful model for studies of embryonic development. It is an especially important model for spiralian development, and for studies of asymmetric cell division. The embryos are amenable to classic embryological manipulation techniques as well as a growing number of molecular approaches. Ilyanassa is also an important model for studies of metamorphosis, the ecology of parasitism, the effects of environmental contaminants on morphology and sexual function, and comparative neurobiology. Ilyanassa is host to several species of parasitic trematode worms, so care must be taken to avoid contamination of Ilyanassa genomic DNA with that of the parasites. The easiest way to avoid this contamination is to isolate DNA from veliger larvae, which are not parasitized. This also avoids other problems that can be encountered when isolating DNA from adult mollusc tissues, such as the presence of large amounts of polysaccharides. This protocol describes the isolation of genomic DNA from Ilyanassa larvae.
海洋腹足纲动物伊利亚那萨螺(Ilyanassa obsoleta)是胚胎发育研究中一个长期且非常有用的模型。它是螺旋动物发育以及不对称细胞分裂研究的一个特别重要的模型。这些胚胎适用于经典的胚胎学操作技术以及越来越多的分子方法。伊利亚那萨螺也是变态发育、寄生生态学、环境污染物对形态和性功能的影响以及比较神经生物学研究的重要模型。伊利亚那萨螺是几种寄生吸虫的宿主,因此必须注意避免伊利亚那萨螺基因组DNA被寄生虫的DNA污染。避免这种污染的最简单方法是从未被寄生的面盘幼虫中分离DNA。这也避免了从成年软体动物组织中分离DNA时可能遇到的其他问题,例如存在大量多糖。本方案描述了从伊利亚那萨螺幼虫中分离基因组DNA的方法。