Exponential amplification of recombinant-RNA hybridization probes
Por:
Lizardi P.M., Guerra C.E., Lomeli H., Tussie-Luna I., Kramer F.R.
Publicada:
1 ene 1988
Resumen:
We have synthesized recombinant RNA molecules that function both as hybridization probes and as templates for exponential amplification by Qð??? replicase. Each recombinant consists of a sequence specific for the protozoan parasite, Plasmodium, falciparum, embedded within the sequence of MDV-1 RNA, which is a natural template for Qð??? replicase. The probe sequence was inserted within a hairpin loop that occurs on the exterior of MDV-1 RNA. The recombinant RNAs hybridize specifically to complementary DNA, despite topological constraints on the probe domain, are replicated at the same rate as MDV-1 RNA, despite their additional length, and are able to serve as templates for the synthesis of a large number of RNA copies. A Qð??? replicase reaction initiated with only 0.14 femtograms of recombinant RNA (1, 000 molecules) can produce 129 nanograms of recombinant RNA product in 30 minutes. This represents a one-billion fold amplification. Our results demonstrate the feasibility of employing exponentially replicatable RNAs in bioassays, where they would serve the dual role of specific probe and amplifiable reporter. © 1988 Nature Publishing Group.
Filiaciones:
Lizardi P.M.:
Centro de Investigacion sobre Ingeniería Genética y Biotecnología, Universidad Nacional Autónoma de México, Apartado Postal 510-3, Cuernavaca, 62270, United States
Guerra C.E.:
Centro de Investigacion sobre Ingeniería Genética y Biotecnología, Universidad Nacional Autónoma de México, Apartado Postal 510-3, Cuernavaca, 62270, United States
Lomeli H.:
Centro de Investigacion sobre Ingeniería Genética y Biotecnología, Universidad Nacional Autónoma de México, Apartado Postal 510-3, Cuernavaca, 62270, United States
Tussie-Luna I.:
Centro de Investigacion sobre Ingeniería Genética y Biotecnología, Universidad Nacional Autónoma de México, Apartado Postal 510-3, Cuernavaca, 62270, United States
Kramer F.R.:
Centro de Investigacion sobre Ingeniería Genética y Biotecnología, Universidad Nacional Autónoma de México, Apartado Postal 510-3, Cuernavaca, 62270, United States
Department of Molecular Genetics, Public Health Research Institute, 455 First Avenue, New York, NY, 10016, United States
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