Highly sensitive genotyping using artificial riboregulator system.
A molecular-beacon-type riboregulator (mRNA) was applied to multiply catalytic gene sensing. It consists of a reporter gene for firefly protein luciferase and, upstream thereof, a regulator hairpin domain composed of an RBS/anti-RBS stem (RBS = ribosome binding site) and a loop which is complementary to the target. The sensing of target gene, using an unmodified RNA or even dsDNA as a probe with a chemiluminescence output, was demonstrated with a sensitivity at < or = 50 fmol of the target and a single nucleotide resolution.
Narita
Atsushi
A
Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto university, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.
Ogawa
Kazumasa
K
Sando
Shinsuke
S
Aoyama
Yasuhiro
Y
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D017930:Genes, Reporter; D005838:Genotype; D049409:Luciferases, Firefly / Q000235:genetics; D049408:Luminescent Agents / Q000737:chemistry*; D008163:Luminescent Measurements; D015345:Oligonucleotide Probes / Q000737:chemistry; D014176:Protein Biosynthesis; D015347:RNA Probes / Q000737:chemistry*; D058727:RNA, Small Untranslated; D022661:RNA, Untranslated / Q000378:metabolism; D019713:Receptors, CCR5 / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*; D012808:Sigma Factor / Q000096:biosynthesis / Q000235:genetics
D001426:Bacterial Proteins; C405431:DsrA RNA, E coli; D049408:Luminescent Agents; D015345:Oligonucleotide Probes; D015347:RNA Probes; D058727:RNA, Small Untranslated; D022661:RNA, Untranslated; D019713:Receptors, CCR5; D012808:Sigma Factor; C069567:sigma factor KatF protein, Bacteria; D049409:Luciferases, Firefly