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Probing mRNA structure and sRNA-mRNA interactions in bacteria using enzymes and lead(II)
- Source :
- Methods in Molecular Biology, Methods in Molecular Biology, Humana Press/Springer Imprint, 2009, 540, pp.215-32. ⟨10.1007/978-1-59745-558-9_16⟩, Methods in Molecular Biology (Clifton then Totova), Methods in Molecular Biology (Clifton then Totova), Humana Press (Springer Imprint), 2009, 540, pp.215-32. 〈10.1007/978-1-59745-558-9_16〉
- Publication Year :
- 2009
- Publisher :
- HAL CCSD, 2009.
-
Abstract
- International audience; Enzymatic probing and lead(II)-induced cleavages have been developed to study the secondary structure of RNA molecules either free or engaged in complex with different ligands. Using a combination of probes with different specificities (unpaired vs. paired regions), it is possible to get information on the accessibility of each nucleotide, on the binding site of a ligand (noncoding RNAs, protein, metabolites), and on RNA conformational changes that accompanied ligand binding or environmental conditions (temperature, pH, ions, etc.). The detection of the cleavages can be conducted by two different ways, which are chosen according to the length of the studied RNA. The first method uses end-labeled RNA molecules and the second one involves primer extension by reverse transcriptase. We provide here an experimental procedure that was designed to map the structure of mRNA and mRNA-sRNA interaction in vitro.
- Subjects :
- Staphylococcus aureus
RNA, Untranslated
MESH : RNA, Messenger
MESH : RNA, Untranslated
MESH : Molecular Sequence Data
Molecular Sequence Data
MESH : Nucleic Acid Conformation
MESH: Enzymes
MESH : Enzymes
[ SDV.BBM.BM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology
[SDV.BC]Life Sciences [q-bio]/Cellular Biology
Chemical Fractionation
MESH: Base Sequence
MESH: RNA, Untranslated
MESH: Chemical Fractionation
MESH : Hydrolysis
MESH: RNA
MESH : RNA
MESH: Staphylococcus aureus
RNA, Messenger
Molecular Biology
MESH: RNA, Messenger
MESH: Molecular Sequence Data
Base Sequence
MESH : Chemical Fractionation
[ SDV.BC ] Life Sciences [q-bio]/Cellular Biology
Hydrolysis
MESH : Staphylococcus aureus
MESH: Molecular Biology
[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology
MESH : Molecular Biology
MESH : RNA, Bacterial
Enzymes
RNA, Bacterial
MESH: Nucleic Acid Conformation
Lead
Nucleic Acid Conformation
RNA
MESH : Base Sequence
MESH : Lead
MESH: RNA, Bacterial
MESH: Hydrolysis
MESH: Lead
Subjects
Details
- Language :
- English
- ISSN :
- 10643745 and 19406029
- Database :
- OpenAIRE
- Journal :
- Methods in Molecular Biology, Methods in Molecular Biology, Humana Press/Springer Imprint, 2009, 540, pp.215-32. ⟨10.1007/978-1-59745-558-9_16⟩, Methods in Molecular Biology (Clifton then Totova), Methods in Molecular Biology (Clifton then Totova), Humana Press (Springer Imprint), 2009, 540, pp.215-32. 〈10.1007/978-1-59745-558-9_16〉
- Accession number :
- edsair.pmid.dedup....79c7500e56d1b5045d515ccb7a8e888e