High Processivity Polymerases
AssigneeWilliam Marsh Rice University
InventorsYousif Shamoo, Siyang Sun
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Abstract
Chimeric proteins comprising a sequence nonspecific single-stranded nucleic-acid-binding domain joined to a catalytic nucleic-acid-modifying domain are provided. Methods comprising contacting a nucleic acid molecule with a chimeric protein, as well as systems comprising a nucleic acid molecule, a chimeric protein, and an aqueous solution are also provided. The joining of sequence nonspecific single-stranded nucleic-acid-binding domain and a catalytic nucleic-acid-modifying domain in chimeric proteins, among other things, may prevent the separation of the two domains due to their weak association and thereby enhances processivity while maintaining fidelity.
Claims21
A chimeric protein comprising a sequence nonspecific single-stranded nucleic-acid-binding domain joined to a catalytic nucleic-acid-modifying domain.
A method comprising contacting a nucleic acid molecule with a chimeric protein having at least two heterologous domains, wherein a first domain that is a sequence-non-specific single-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 single-stranded nucleic-acid-binding domain stabilizes the formation of a single-stranded nucleic acid compared to an otherwise identical protein not having the sequence-non-specific single-stranded nucleic-acid-binding domain joined to it.
A system comprising a nucleic acid molecule and a chimeric protein having at least two heterologous domains, wherein a first domain that is a sequence non-specific single-stranded nucleic-acid-binding domain is joined to a second domain that is a catalytic nucleic-acid-modifying domain;
- and an aqueous solution that permits the binding domain to bind to the nucleic acid molecule and that permits the chimeric protein to function in a catalytic manner to modify the nucleic acid molecule.
Dependencies are inferred from the claim text.