کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
8340236 1541221 2016 10 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Single-molecule pull-down for investigating protein-nucleic acid interactions
ترجمه فارسی عنوان
تک مولکول کشف کردن برای بررسی اثرات متقابل پروتئین-اسید نوکلئیک
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
چکیده انگلیسی
The genome and transcriptome are constantly modified by proteins in the cell. Recent advances in single-molecule techniques allow for high spatial and temporal observations of these interactions between proteins and nucleic acids. However, due to the difficulty of obtaining functional protein complexes, it remains challenging to study the interactions between macromolecular protein complexes and nucleic acids. Here, we combined single-molecule fluorescence with various protein complex pull-down techniques to determine the function and stoichiometry of ribonucleoprotein complexes. Through the use of three examples of protein complexes from eukaryotic cells (Drosha, Dicer, and TUT4 protein complexes), we provide step-by-step guidance for using novel single-molecule techniques. Our single-molecule methods provide sub-second and nanometer resolution and can be applied to other nucleoprotein complexes that are essential for cellular processes.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Methods - Volume 105, 1 August 2016, Pages 99-108
نویسندگان
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