Nucleic acid modifying enzymes
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.
1. A polymerase comprising two joined heterologous domains:
a sequence non-specific double-stranded nucleic acid binding domain, wherein the sequence non-specific double-stranded nucleic acid binding domain is a HMF family Archaeal histone; and
a DNA polymerase domain that has DNA polymerization activity,
wherein the sequence non-specific double-stranded nucleic acid binding domain and the DNA polymerase domain are covalently linked and the presence of the sequence non-specific double-stranded nucleic acid binding domain enhances the processivity of the polymerase domain compared to an identical protein that does not have the sequence non-specific double-stranded nucleic acid binding domain joined thereto.
2. A method of enhancing the processivity of a polymerase, the method comprising joining a heterologous sequence non-specific double stranded nucleic acid binding domain to a polymerase domain, wherein the sequence non-specific double-stranded nucleic acid binding domain is a HMF family Archaeal histone; and thereby enhancing the processivity of the polymerase compared to an identical protein that does not have the sequence non-specific double-stranded nucleic acid binding domain joined thereto.
3. A method of polymerization to form double-stranded nucleic acid, comprising the steps of:
a) contacting a template nucleic acid and a primer with a polymerase of claim 1 ; in a reaction mixture suitable for the polymerase to extend the primer; and
b) allowing polymerization to occur, thereby forming double-stranded nucleic acid.