title: A minimalist biosensor: Quantitation of cyclic di-GMP using the conformational change of a riboswitch aptamer.
abstract: Cyclic di-GMP (c-di-GMP) is a second messenger that is important in regulating bacterial physiology and behavior, including motility and virulence. Many questions remain about the role and regulation of this signaling molecule, but current methods of detection are limited by either modest sensitivity or requirements for extensive sample purification. We have taken advantage of a natural, high affinity receptor of c-di-GMP, the Vc2 riboswitch aptamer, to develop a sensitive and rapid electrophoretic mobility shift assay (EMSA) for c-di-GMP quantitation that required minimal engineering of the RNA.
authors.affiliation: a Department of Chemistry ; University of California, Berkeley ; Berkeley , CA USA.
authors.forename: Colleen A
authors.initials: CA
authors.lastname: Kellenberger
authors.affiliation: a Department of Chemistry ; University of California, Berkeley ; Berkeley , CA USA.
authors.forename: Jade
authors.initials: J
authors.lastname: Sales-Lee
authors.affiliation: b Graduate Program in Bioengineering; University of California, Berkeley ; Berkeley , CA USA.
authors.forename: Yuchen
authors.initials: Y
authors.lastname: Pan
authors.affiliation: a Department of Chemistry ; University of California, Berkeley ; Berkeley , CA USA.
authors.forename: Madalee M
authors.initials: MM
authors.lastname: Gassaway
authors.affiliation: b Graduate Program in Bioengineering; University of California, Berkeley ; Berkeley , CA USA.
authors.forename: Amy E
authors.initials: AE
authors.lastname: Herr
authors.affiliation: a Department of Chemistry ; University of California, Berkeley ; Berkeley , CA USA.|c Department of Molecular & Cell Biology ; University of California, Berkeley ; Berkeley , CA USA.
authors.forename: Ming C
authors.initials: MC
authors.lastname: Hammond
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D015374:Biosensing Techniques; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000737:chemistry; D024202:Electrophoretic Mobility Shift Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D009690:Nucleic Acid Conformation; D058928:Riboswitch / Q000235:genetics; D015290:Second Messenger Systems; D015398:Signal Transduction
chemical_list: D052157:Aptamers, Nucleotide; D058928:Riboswitch; C062025:bis(3',5')-cyclic diguanylic acid; D006152:Cyclic GMP
keywords: bacterial signaling; cyclic AMP-GMP; cyclic dinucleotides; ligand binding; microfluidic mobility shift assay; native EMSA