Single and dual drug selection for transgenes following bombardment of Caenorhabditis species.
Single and dual drug selection for transgenes following bombardment of Caenorhabditis species.
复制标题
轰击秀丽隐杆线虫后转基因的单药和双药选择。
DOI：
10.1016/j.ymeth.2014.05.004
复制
发表时间：
2014
期刊：
影响因子：
4.8
通讯作者：
Ben Lehner
中科院分区：
文献类型：
--
作者：
J. Semple;Ben Lehner
使用药物和耐药基因是选择转基因存在的有效方法。与需要基因突变互补的方法不同，该系统可以在任何遗传背景下使用。药物选择不需要大量的操作或昂贵的设备，但速度非常快，并且可以达到极高的效率，从数百万条非转基因蠕虫中选择出少量的转基因蠕虫。通过微粒轰击将整合转基因引入秀丽隐杆线虫正是这样一个挑战。在这里，我们详细描述了我们为嘌呤霉素和 G418 在液体中进行双药选择而开发的方案，该方案在多种线虫物种中效果良好。我们还表明，仅使用嘌呤霉素或仅使用 G418 的单一药物选择在 C 中是有效的。线虫。越来越多的药物选择标记已适应 C.线虫是我们可用的遗传工具包的重要补充。
The use of drugs and drug resistance genes is a powerful method to select for the presence of a transgene. Unlike methods that require the complementation of a genetic mutation, this system can be used on any genetic background. Drug selection does not require extensive manipulation or costly equipment, yet it is very rapid and can achieve extremely high efficiency, selecting a small number of transgenic worms from among millions of non-transgenic worms. Introducing integrated transgenes intoCaenorhabditis elegansby microparticle bombardment represents just such a challenge. Here we describe in detail the protocol we have developed for dual-drug selection in liquid with puromycin and G418 which works well in a variety ofCaenorhabditisspecies. We also show that single drug selection with only puromycin or only G418 is effective inC. elegans. The growing number of drug selection markers that have been adapted toC. elegansare an important addition to the genetic toolkit at our disposal.