In Vitro/In Vivo Production of tRNA for X-Ray Studies

Clément Dégut; Alexandre Monod; Franck Brachet; Thibaut Crépin; Carine Tisné

doi:10.1007/978-1-4939-2763-0_4

In Vitro/In Vivo Production of tRNA for X-Ray Studies

Methods Mol Biol. 2016:1320:37-57. doi: 10.1007/978-1-4939-2763-0_4.

Authors

Clément Dégut¹, Alexandre Monod, Franck Brachet, Thibaut Crépin, Carine Tisné

Affiliation

¹ Laboratoire de Cristallographie et RMN Biologiques, CNRS, Paris Sorbonne Cité, 4 avenue de l'Observatoire, Paris, 75006, France.

PMID: 26227036
DOI: 10.1007/978-1-4939-2763-0_4

Abstract

tRNAs occupy a central role in the cellular life, and they are involved in a broad range of biological processes that relies on their interaction with proteins and RNA. Crystallization and structure resolution of tRNA or/and tRNA/partner complexes can yield in valuable information on structural organizations of key elements of cellular machinery. However, crystallization of RNA, is often challenging. Here we review two methods to produce and purify tRNA in quantity and quality to perform X-ray studies.

MeSH terms

Base Sequence
Chromatography
Crystallization
Crystallography, X-Ray / methods*
DNA-Directed RNA Polymerases / chemistry
Escherichia coli / enzymology
Hydrogen-Ion Concentration
Magnetic Resonance Spectroscopy
Molecular Sequence Data
Nucleic Acid Conformation
Plasmids
RNA, Transfer / chemistry*
Reproducibility of Results
Ribonucleases / chemistry
Transcription, Genetic
Viral Proteins / chemistry
X-Rays

Substances

Viral Proteins
RNA, Transfer
bacteriophage T7 RNA polymerase
DNA-Directed RNA Polymerases
Ribonucleases