Rna-dependent rna polymerase functioning preferably on rna...

Details Rna-dependent rna polymerase functioning preferably on rna... Rna-dependent rna polymerase functioning preferably on rna...

C - Chemistry, Metallurgy

12

N

C12N 9/12 (2006.01) C12N 15/54 (2006.01) C12N 15/63 (2006.01) C12P 19/34 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2286343

Disclosed is a process for transcribing RNA using a nucleotide reagent as the promoter. Such a reagent enables any type of RNA to be transcribed without sequence specification and without protein cofactors, by means of an RNA polymerase that is known to be DNA-dependent such as the RNA polymerase of the phage T7, or by means of new, mutated RNA polymerase with the ability to synthesize a transcription product of a polynucleotide matrix with a higher yield when the matrix is RNA than when said matrix is DNA. This type of RNA polymerase can be obtained by effecting mutations on a coding gene for a wild-type RNA polymerase, and then by selecting the mutated RNA polymerase with said ability. The invention can be applied notably to the detection, synthesis or quantification of RNA.

Procédé de transcription d'ARN polymérase à l'aide d'un réactif nucléotidique jouant le rôle de promoteur. Un tel réactif permet de transcrire un ARN quelconque, sans spécificité de séquence, et en l'absence de cofacteurs protéiques, à l'aide d'ARN polymérases connues comme étant ADN dépendantes, telles que l'ARN polymérase du phage T7, ou encore à l'aide de nouvelles ARN polymérases mutées ayant la propriété de synthétiser un produit de transcription d'une matrice polynucléotidique avec un meilleur rendement lorsque la matrice est de l'ARN que lorsque la matrice est de l'ADN. Une telle ARN polymérase peut être obtenue en effectuant des mutations sur un gène codant pour une ARN polymérase de type sauvage et en sélectionnant les ARN polymérases mutées ayant ladite propriété. Application notamment à la détection, la synthèse ou la quantification d'ARN.

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