Coexpression and Copurification of RNA-Protein Complexes in Escherichia coli.
Fiche publication
Date publication
janvier 2021
Journal
Methods in molecular biology (Clifton, N.J.)
Auteurs
Membres identifiés du Cancéropôle Est :
Pr DARDEL Frédéric
Tous les auteurs :
El Khouri M, Catala M, Seijo B, Chabal J, Dardel F, Tisné C, Ponchon L
Lien Pubmed
Résumé
For structural, biochemical, or pharmacological studies, it is required to have pure RNA in large quantities. We previously devised a generic approach that allows for efficient in vivo expression of recombinant RNA in Escherichia coli. We have extended the "tRNA scaffold" method to RNA-protein coexpression 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.
Mots clés
Expression plasmid, Protein expression, RNA purification, Recombinant RNA, tRNA scaffold
Référence
Methods Mol Biol. 2021 ;2323:67-73