大肠杆菌中RNA-蛋白质复合物的表达与纯化

Expression and Purification of RNA-Protein Complexes in Escherichia coli.

作者信息

El Khouri Margueritte, Catala Marjorie, Seijo Bili, Chabal Johana, Tisné Carine, Dardel Frédéric, Ponchon Luc

机构信息

Center for Molecular OncologyBarts Cancer Institute, Queen Mary University of London, London, UK.

出版信息

Methods Mol Biol. 2015;1316:25-31. doi: 10.1007/978-1-4939-2730-2_3.

DOI:10.1007/978-1-4939-2730-2_3

PMID:25967050

Abstract

For structural, biochemical or pharmacological studies, it is required to have pure RNA in large quantities. We previously devised a generic approach that allows efficient in vivo expression of recombinant RNA in Escherichia coli. We have extended the "tRNA scaffold" method to RNA/protein co-expression in order to express and purify RNA by affinity in native condition. As a proof-of-concept, we present the expression and the purification of the AtRNA-mala in complex with the MS2 coat protein.

摘要

对于结构、生化或药理学研究而言，需要大量的纯RNA。我们之前设计了一种通用方法，可使重组RNA在大肠杆菌中高效地在体内表达。我们已将“tRNA支架”方法扩展至RNA/蛋白质共表达，以便在天然条件下通过亲和作用表达和纯化RNA。作为概念验证，我们展示了与MS2外壳蛋白复合的AtRNA-mala的表达和纯化。