High fidelity polymerases and uses thereof

Details High fidelity polymerases and uses thereof High fidelity polymerases and uses thereof

C - Chemistry, Metallurgy

07

H

C07H 21/04 (2006.01) C12N 9/12 (2006.01) C12P 19/34 (2006.01)

Patent

CA 2302393

The present invention relates to a DNA and RNA polymerases which have increased fidelity (or reduced misincorporation rate). In particular, the invention relates to a method of making such polymerases by modifying or mutating the nucleotide binding domain of the polymerase (e.g., the O-helix). The invention also relates to DNA molecules containing the genes encoding the polymerases of the invention, to host cells containing such DNA molecules and to methods to make the polymerases using the host cells. The polymerases are particularly suited for nucleic acid synthesis, sequencing, amplification and cDNA synthesis.

L'invention concerne des polymérases ADN et ARN présentant une fidélité accrue (ou à taux d'incorporation erroné réduit). Elle concerne, en particulier, un procédé de préparation de telles polymérases par modification ou mutation du domaine de liaison nucléotidique de la polymérase (par exemple, hélice O). L'invention concerne en outre des molécules d'ADN renfermant des gènes codant pour les polymérases de l'invention, des cellules hôtes contenant de telles molécules d'ADN, ainsi que des procédés de préparation des polymérases au moyen de ces cellules hôtes. Les polymérases conviennent en particulier pour la synthèse, le séquençage, l'amplification d'acides nucléiques et la synthèse d'ADN complémentaire.

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