生物发光
膜
细胞器
细胞生物学
人口
化学
转染
生物物理学
生物
生物化学
基因
医学
环境卫生
作者
Jeannette Tóth,Gergő Gulyás,László Hunyady,Péter Várnai
出处
期刊:Methods in molecular biology
日期:2019-01-01
卷期号:: 23-34
被引量:5
标识
DOI:10.1007/978-1-4939-9136-5_3
摘要
There are several difficulties to face when investigating the role of phosphoinositides. Although they are present in most organelles, their concentration is very low, sometimes undetectable with the available methods; moreover, their level can quickly change upon several external stimuli. Here we introduce a newly improved lipid sensor tool-set based on the balanced expression of luciferase-fused phosphoinositide recognizing protein domains and a Venus protein targeted to the plasma membrane, allowing us to perform Bioluminescence Resonance Energy Transfer (BRET) measurements that reflect phosphoinositide changes in a population of transiently transfected cells. This method is highly sensitive, specific, and capable of semiquantitative characterization of plasma membrane phosphoinositide changes with high temporal resolution.
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