Aix Marseille University, CNRS, IBDM, Turing Centre for Living Systems, Marseille, France.
Methods Mol Biol. 2022;2540:251-268. doi: 10.1007/978-1-0716-2541-5_12.
Proteins are typically not expressed homogeneously in all cells of a complex organism. Within cells, proteins can dynamically change locations, be transported to their destinations, or be degraded upon external signals. Thus, revealing the cellular and subcellular localizations as well as the temporal dynamics of a protein provides important insights into the possible function of the studied protein. Tagging a protein of interest with a genetically encoded fluorophore enables us to follow its expression dynamics in the living organism. Here, we summarize the genetic resources available for tagged Drosophila proteins that assist in studying protein expression and dynamics. We also review the various techniques used in the past and at present to tag a protein of interest with a genetically encoded fluorophore. Comparing the pros and cons of the various techniques guides the reader to judge the suitable applications possible with these tagged proteins in Drosophila.
蛋白质通常不会在复杂生物体的所有细胞中均匀表达。在细胞内,蛋白质可以动态改变位置,被运输到目的地,或者在外部信号作用下被降解。因此,揭示蛋白质的细胞和亚细胞定位以及时空动态为研究蛋白质的可能功能提供了重要的见解。用遗传编码的荧光蛋白标记感兴趣的蛋白质可以使我们在活体生物中追踪其表达动力学。在这里,我们总结了可用于标记果蝇蛋白的遗传资源,这些资源有助于研究蛋白质的表达和动态。我们还回顾了过去和现在用于用遗传编码荧光蛋白标记感兴趣的蛋白质的各种技术。比较各种技术的优缺点,可以指导读者判断这些标记蛋白在果蝇中可能的适用应用。