Real time in vitro analysis of transcription by RNA polymerase on immobilized DNA fibres.
We have used surface plasmon resonance (SPR) to follow variations in the concentrations of binary complexes as RNA polymerase moves into a transcriptionally competent initiation complex with immobilized DNA fibres containing promoter sequences. The use of SPR to follow complex binding phenomena is described. We have also followed the changes in the mass of initiation complexes following addition of the nucleotide triphosphates prerequisite for transcription on the immobilized template. These signals are interpreted in terms of the escape of RNA polymerase into elongation mode and the subsequent synthesis of nascent RNA molecules.
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MESH: DNA; MESH: DNA-Directed RNA Polymerases; MESH: Gene Expression Profiling; MESH: Promoter Regions (Genetics); MESH: Surface Plasmon Resonance; MESH: Templates, Genetic; MESH: Time Factors; MESH: Transcription, Genetic; [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology
Journal articles; Life Sciences [q-bio]; Ecole Normale Supérieure de Cachan; Laboratoire de Biotechnologie et Pharmacologie génétique Appliquée; CNRS - Centre national de la recherche scientifique; Institut Pasteur
ISSN: 0952-3499; EISSN: 1099-1352; Journal of Molecular Recognition; https://hal.archives-ouvertes.fr/hal-00277557; Journal of Molecular Recognition, Wiley, 1999, 12 (5), pp.322-7. ⟨10.1002/(SICI)1099-1352(199909/10)12:53.0.CO;2-W⟩