Gunadi Andika, Dean Eric A, Finer John J
Department of Horticulture and Crop Science, The Ohio State University, Ohio Agricultural Research and Development Center, Wooster, OH, USA.
Pairwise, Research Triangle Park, NC, USA.
Methods Mol Biol. 2019;1864:67-79. doi: 10.1007/978-1-4939-8778-8_5.
Transient transformation or transient expression results in rapid and fleeting gene expression. This approach is often used as a first-tier screening tool for evaluation of components that affect gene expression. Here, we describe the use of particle bombardment of lima bean cotyledons with constructs containing the green fluorescent protein (gfp) coding region for evaluation of promoter components that influence gene expression. Although this approach is conceptually quite simple, this lima bean transient expression system may not work well, if our methods and notes are not carefully read and followed. Our laboratory has successfully optimized this method over the past 10 years, resulting in a transient expression system, which works like no other that we have seen.
瞬时转化或瞬时表达会导致基因快速短暂表达。这种方法常被用作评估影响基因表达的元件的一级筛选工具。在此，我们描述了用含有绿色荧光蛋白（gfp）编码区的构建体对利马豆子叶进行粒子轰击，以评估影响基因表达的启动子元件。尽管这种方法在概念上相当简单，但如果不仔细阅读并遵循我们的方法和注意事项，这个利马豆瞬时表达系统可能效果不佳。在过去10年里，我们实验室成功优化了此方法，得到了一个与我们见过的其他系统都不同的瞬时表达系统。
