Coexpression and Copurification of RNA-Protein Complexes in Escherichia coli.
Ampicillin
/ pharmacology
Aptamers, Nucleotide
/ chemistry
Capsid
Chloramphenicol
/ pharmacology
Chromatography, Affinity
/ methods
Cloning, Molecular
/ methods
Computer Simulation
Drug Resistance, Microbial
/ genetics
Electrophoresis, Polyacrylamide Gel
/ methods
Escherichia coli
/ chemistry
Escherichia coli Proteins
/ biosynthesis
Levivirus
/ genetics
Models, Molecular
Nucleic Acid Conformation
Operator Regions, Genetic
Plasmids
/ genetics
RNA
/ biosynthesis
RNA, Bacterial
/ genetics
RNA, Viral
/ genetics
RNA-Binding Proteins
/ biosynthesis
Expression plasmid
Protein expression
RNA purification
Recombinant RNA
tRNA scaffold
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2021
2021
Historique:
entrez:
4
6
2021
pubmed:
5
6
2021
medline:
20
8
2021
Statut:
ppublish
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.
Identifiants
pubmed: 34086274
doi: 10.1007/978-1-0716-1499-0_6
doi:
Substances chimiques
Aptamers, Nucleotide
0
Escherichia coli Proteins
0
RNA, Bacterial
0
RNA, Viral
0
RNA, recombinant
0
RNA-Binding Proteins
0
malachite green RNA aptamer
0
RNA
63231-63-0
Chloramphenicol
66974FR9Q1
Ampicillin
7C782967RD
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
67-73Références
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