Method for cloning and expression of Rhodothermus Obamensis DNA polymerase I large fragment in E. coli
AssigneeNew England Biolabs, Inc.
InventorShuang-yong Xu
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Abstract
The present invention provides a novel thermostable DNA polymerase I obtainable from Rhodothermus obamensis, which possesses 3′-5′ exonuclease activity and has a half-life of about 35 minutes at 94° C. This polymerase also contains a tyrosine residue in the ribosome binding site which improves incorporation of dideoxyribonucleic acids. Also provided are isolated DNA and vectors encoding this polymerase, as well as its large fragment, and methods for producing recombinant enzyme using the same.
Claims6
A substantially pure thermostable DNA polymerase I obtainable from Rhodothermus obamensis (JCM 9785), wherein said polymerase possesses 3′-5′ exonuclease activity and has a half-life of about 35 minutes at 94° C.
A recombinant R. obamensis DNA polymerase I, wherein said polymerase I has a molecular weight of about 104 kDa, possesses 3′-5′ exonuclease activity and has a half-life of about 35 minutes at 94° C.
A recombinant R. obamensis DNA polymerase I large fragment, wherein said polymerase I large fragment has a molecular weight of about 71 kDa, possesses 3′-5′ exonuclease activity and has a half-life of about 35 minutes at 94° C.
Dependencies are inferred from the claim text.