芽殖酵母
核孔
酵母
细胞生物学
活体细胞成像
核成像
细胞
生物
生物物理学
计算生物学
酿酒酵母
化学
纳米技术
遗传学
材料科学
细胞质
医学
核医学
作者
Abdolaziz Rastegar Lari,Farzin Farzam,Pierre Bensidoun,Marlene Oeffinger,Daniel Zenklusen,David Grünwald,Ben Montpetit
出处
期刊:Methods in molecular biology
日期:2019-01-01
卷期号:: 131-150
被引量:3
标识
DOI:10.1007/978-1-4939-9674-2_9
摘要
Single-molecule resolution imaging has become an important tool in the study of cell biology. Aptamer-based approaches (e.g., MS2 and PP7) allow for detection of single RNA molecules in living cells and have been used to study various aspects of mRNA metabolism, including mRNP nuclear export. Here we outline an imaging protocol for the study of interactions between mRNPs and nuclear pore complexes (NPCs) in the yeast S. cerevisiae, including mRNP export. We describe in detail the steps that allow for high-resolution live-cell mRNP imaging and measurement of mRNP interactions with NPCs using simultaneous two-color imaging. Our protocol discusses yeast strain construction, choice of marker proteins to label the nuclear pore complex, as well as imaging conditions that allow high signal-to-noise data acquisition. Moreover, we describe various aspects of postacquisition image analysis for single molecule tracking and image registration allowing for the characterization of mRNP–NPC interactions.
科研通智能强力驱动
Strongly Powered by AbleSci AI