Saka Yasushi, Hagemann Anja I, Smith James C
Interdisciplinary Research Institute, CNRS USR3078, Institut de Biologie de Lille, 1 rue du Professeur Calmette, Lille Cedex, France.
Methods. 2008 Jul;45(3):192-5. doi: 10.1016/j.ymeth.2008.06.005. Epub 2008 Jun 27.
Bimolecular fluorescence complementation (BiFC) provides a simple and direct way to visualise protein-protein interactions in vivo and in real-time. In this article, we describe methods by which one can implement this approach in embryos of the South African claw-toed frog Xenopus laevis. We have made use of Venus, an improved version of yellow fluorescent protein (YFP), so as to achieve rapid detection of protein interactions. To suppress spontaneous interactions between the N- and C-terminal fragments of Venus, a point mutation (T153M) was introduced into the N-terminal fragment. We have used this reagent to monitor signalling by members of the transforming growth factor type beta family in cells of the Xenopus embryo.
双分子荧光互补（BiFC）提供了一种简单直接的方法，可在体内实时可视化蛋白质-蛋白质相互作用。在本文中，我们描述了在南非爪蟾非洲爪蟾胚胎中实施此方法的步骤。我们利用了黄色荧光蛋白（YFP）的改进版本Venus，以实现对蛋白质相互作用的快速检测。为了抑制Venus的N端和C端片段之间的自发相互作用，在N端片段中引入了一个点突变（T153M）。我们已使用该试剂监测非洲爪蟾胚胎细胞中转化生长因子β家族成员的信号传导。
