Rieckher Matthias, Kourtis Nikos, Pasparaki Angela, Tavernarakis Nektarios
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, Heraklion, Crete, Greece.
Methods Mol Biol. 2009;561:21-39. doi: 10.1007/978-1-60327-019-9_2.
Two efficient strategies have been developed and are widely used for the genetic transformation of the nematode Caenorhabditis elegans, DNA microinjection, and DNA-coated microparticle bombardment. Both methodologies facilitate the delivery of exogenous DNA into the developing oocytes of adult hermaphrodite animals, which then generate transgenic worms among their progeny. Although both approaches share the common underlying principle of introducing foreign DNA into the germline of C. elegans, they offer distinct transformation outcomes. In this chapter, we present DNA microinjection and bombardment methods for transgenesis in C. elegans and provide time-tested procedures for their implementation. We also discuss their relative advantages as well as their limitations and evaluate their potential for a range of applications.
已经开发出两种高效的策略并广泛用于线虫秀丽隐杆线虫的基因转化，即DNA显微注射和DNA包被的微粒轰击。这两种方法都有助于将外源DNA传递到成年雌雄同体动物发育中的卵母细胞中，这些卵母细胞随后在其后代中产生转基因蠕虫。尽管这两种方法都具有将外源DNA引入秀丽隐杆线虫种系的共同基本原理，但它们提供了不同的转化结果。在本章中，我们介绍了秀丽隐杆线虫转基因的DNA显微注射和轰击方法，并提供了经过时间考验的实施程序。我们还讨论了它们的相对优势以及局限性，并评估了它们在一系列应用中的潜力。
