Mutant chimeric dna polymerase

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

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

Patent

CA 2240570

The invention provides mutant, chimeric thermostable DNA polymerase enzymes, which chimeric thermostable DNA polymerase enzymes consist of an N-terminal region derived from the 5'-nuclease domain of a Thermus species DNA polymerase and a C--terminal region derived from the 3' to 5' exonuclease andpolymerase domains of Tma DNA polymerase. These mutant chimeric thermostable DNA polymerase enzymes have improved properties in nucleic acid sequencing reactions. Also provided are nucleic acids encoding said mutant chimeric thermostable DNA polymerase enzymes, vectors comprising said nucleic acids and host cells transformed with said vectors. Also provided are compositions comprising said mutated, chimeric thermostable DNA polymerase enzymes and non-ionic polymeric detergent(s). Futhermore methods for producing the said enzymes and methods and kits for using the said enzymes are provided.

L'invention concerne des enzymes d'ADN polymérase chimérique mutante, lesquelles se composent d'une région N--terminal dérivée du domaine de nucléase 5' d'un ADN polymérase d'espèce Thermus et d'une région C--terminal dérivée des domaines 3 'à 5 'exonucléase andpolymerase d'un ADN polymérase Ttma. Ces enzymes d'ADN polymérase thermostable chimérique mutante présentent des propriétés améliorées dans des réactions de séquençage d'acide nucléique. L'invention concerne également des acides nucléiques codant pour ledit ADN polymérase thermostable chimérique mutante, des vecteurs comprenant lesdits acides nucléiques, et des cellules hôtes transformées par ces vecteurs. L'invention porte également sur des compositions comprenant ledit ADN polymérase thermostable chimérique muté et un ou des détergents de polymère non ionique. En outre, on présente des procédés de production de ces enzymes, des procédés et des nécessaires d'utilisation de ces enzymes.

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