Nucleic acid modifying enzymes
AssigneeMJ Bioworks Incorporated
InventorYan Wang
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Abstract
This invention provides for an improved generation of novel nucleic acid modifying enzymes. The improvement is the fusion of a sequence-non-specific nucleic-acid-binding domain to the enzyme in a manner that enhances the ability of the enzyme to bind and catalytically modify the nucleic acid.
Claims43
A protein consisting of at least two heterologous domains wherein a first domain that is a sequence-non-specific double-stranded nucleic-acid-binding domain is joined to a second domain that is a catalytic nucleic-acid-modifying domain having a processive nature, where the presence of the sequence-non-specific double-stranded nucleic-acid-binding domain enhances the processive nature of the nucleic-acid-modifying domain compared to an identical protein not having a sequence-non-specific nucleic-acid-binding domain joined thereto.
A protein consisting of at least two heterologous domains, wherein a first domain that is a sequence-non-specific double-stranded nucleic-acid-binding domain is joined to a second domain that is a catalytic nucleic-acid-modifying domain, where the presence of the sequence-non-specific nucleic-acid-binding domain stabilizes the double-stranded conformation of a nucleic acid by at least 1° C. compared to an identical protein not having a sequence-non-specific nucleic acid-binding domain joined thereto.
A method of modifying a nucleic acid in an aqueous solution by:
- (i) contacting the nucleic acid with a protein comprising at least two heterologous domains, wherein a first domain that is a sequence-non-specific nucleic-acid-binding domain is joined to a second domain that is a catalytic nucleic-acid-modifying domain having a processive nature, where the sequence non-specific nucleic-acid-binding domain:
- a. binds to double-stranded nucleic acid, and
- b. enhances the processivity of the enzyme compared to an identical enzyme not having the sequence non-specific nucleic-acid-binding domain fused to it, and
- wherein the solution is at a temperature and of a composition that permits the binding domain to bind to the nucleic acid and the enzyme to function in a catalytic manner, and
- (ii) permitting the catalytic domain to modify the nucleic acid in the solution.
A method of modifying a nucleic acid by:
- (i) contacting the nucleic acid with an aqueous solution containing a protein having at least two heterologous domains, wherein a first domain that is a sequence-non-specific double-stranded nucleic acid-binding domain is joined to a second domain that is a catalytic nucleic-acid-modifying domain, where the presence of the sequence-non-specific nucleic-acid-binding domain s stabilizes the double-stranded conformation of a nucleic acid by at least 1° C. compared to an otherwise identical protein not having the sequence-non-specific nucleic-acid-binding domain joined to it;
- and, wherein the solution is at a temperature and of a composition that permits the binding domain to bind to the nucleic acid and the enzyme to function in a catalytic manner;
- and,
- (ii) permitting the catalytic domain to modify the nucleic acid in the solution.
Dependencies are inferred from the claim text.