Sparkes Imogen A, Runions John, Kearns Anne, Hawes Chris
School of Life Sciences, Oxford Brookes University, Oxford OX3 0BP, UK.
Nat Protoc. 2006;1(4):2019-25. doi: 10.1038/nprot.2006.286.
Expression and tracking of fluorescent fusion proteins has revolutionized our understanding of basic concepts in cell biology. The protocol presented here has underpinned much of the in vivo results highlighting the dynamic nature of the plant secretory pathway. Transient transformation of tobacco leaf epidermal cells is a relatively fast technique to assess expression of genes of interest. These cells can be used to generate stable plant lines using a more time-consuming, cell culture technique. Transient expression takes from 2 to 4 days whereas stable lines are generated after approximately 2 to 4 months.
荧光融合蛋白的表达与追踪彻底改变了我们对细胞生物学基本概念的理解。本文介绍的实验方案为许多体内实验结果奠定了基础,这些结果突出了植物分泌途径的动态性质。烟草叶片表皮细胞的瞬时转化是一种相对快速的技术,用于评估感兴趣基因的表达。这些细胞可通过一种更耗时的细胞培养技术用于生成稳定的植物株系。瞬时表达需要2至4天,而稳定株系则在大约2至4个月后产生。