Thermostable dna polymerase from bacillus pallidus

C - Chemistry – Metallurgy – 12 – N

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C12N 15/54 (2006.01) C07K 14/245 (2006.01) C07K 16/40 (2006.01) C12N 5/18 (2006.01) C12N 9/12 (2006.01) C12N 9/96 (2006.01)

Patent

CA 2210506

A novel Type I DNA polymerase derived from Bacillus pallidus is provided. The gene encoding the polymerase has been cloned and expressed. Characterization of the enzyme revealed, in addition to DNA polymerase activity, 5'-3' exonuclease activity, reverse transcriptase activity, strand displacing activity and the ability to initiate polymerization at a nick in double-stranded DNA. In certain constructs the domain encoding the 5'-3' exonuclease activity is deleted to provide an enzyme which is particularly useful for Strand Displacement Amplification. Anti-Bacillus pallidus Polymerase I antibodies and methods of using the antibodies are also provided.

Une nouvelle ADN-polymérase de type I dérivée de Bacillus pallidus est offerte. Le gène codant la polymérase a été cloné et exprimé. La caractérisation de l'enzyme a révélé l'existence, en plus de l'activité ADN-polymérase, d'une activité 5'-3' exonucléase, d'une activité de transcriptase inverse, d'une activité de déplacement des brins ainsi que la capacité d'amorcer la polymérisation dans une coupure simple brin dans l'ADN bicaténaire. Dans certains construits, le domaine codant l'activité 5'-3' exonucléase est délété pour donner une enzyme qui est particulièrement utile pour l'Amplification par déplacement de brin (Strand Displacement Amplification). Des anticorps dirigés contre la polymérase I de Bacillus pallidus et des méthodes pour utiliser ces anticorps sont également offerts.

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