Mutational Analysis of sRNA-mRNA Base Pairing by Electrophoretic Mobility Shift Assay

Publikation: Kapitel i bog/rapport/konference-proceedingKapitel i bogForskning

Abstrakt

Small regulatory RNAs (sRNAs) in bacteria often act by base pairing to mRNAs. Direct interactions between an sRNA and its target mRNA can be investigated by electrophoretic mobility shift assay. In this assay, regions engaged in base pairing are analyzed by introducing mutations in one of the RNAs that prevent sRNA–mRNA complex formation, followed by the introduction of complementary mutations in its partner RNA that restore base pairing. Here, we describe the design of a mutational strategy used to analyze the base pairing between two CU-rich regions of the sRNA Rli22 and the AG-rich Shine-Dalgarno region of the mRNA oppA in Listeria monocytogenes. The protocol can be employed for mutational studies of base pairing between any sRNA and its mRNA target(s).

OriginalsprogEngelsk
TitelBacterial Regulatory RNA : Methods and Protocols
RedaktørerVéronique Arluison, Claudio Valverde
ForlagHumana Press
Publikationsdato2018
Sider165-176
ISBN (Trykt)978-1-4939-7633-1
ISBN (Elektronisk)978-1-4939-7634-8
DOI
StatusUdgivet - 2018
NavnMethods in Molecular Biology
Vol/bind1737
ISSN1064-3745

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  • Citationsformater

    Lillebæk, E. M. S., & Kallipolitis, B. H. (2018). Mutational Analysis of sRNA-mRNA Base Pairing by Electrophoretic Mobility Shift Assay. I V. Arluison, & C. Valverde (red.), Bacterial Regulatory RNA: Methods and Protocols (s. 165-176). Humana Press. Methods in Molecular Biology, Bind. 1737 https://doi.org/10.1007/978-1-4939-7634-8_10