Hyperactive, non-phosphorylated, mutant transposases of mariner mobile genetic elements
AssigneesCentre National de la Recherche Scientifique (CNRS), Universite Francois Rabelais De Tours
InventorsYves Bigot, Corinne Auge-Gouillou, Marie-Hélène Hamelin, Benjamin Brillet
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Abstract
The invention relates to hyperactive, non-phosphorylated, mutant transposases of mariner mobile genetic elements. The invention also relates to recombinant nucleotide sequences encoding such transposases. The invention further relates to a method of producing said transposases and to the use thereof for in vitro or in vivo transposition.
Claims19
A purified mutant hyperactive transposase of a mariner mobile genetic element (MGE), having at least one mutation by conservative substitution of at least one phosphorylatable residue in at least one phosphorylation site, said conservative substitution making said phosphorylation site non-phosphorylatable in vivo, said phosphorylatable residue being selected from the group consisting of:
- the following residues in sequence SEQ ID NO:
- 2:
- T11;
- T24;
- S28;
- T88;
- S99;
- S104;
- T135;
- S147;
- T154;
- S170;
- T181;
- S200;
- T216;
- T255;
- and S305, and
- corresponding residues in the sequence of a mariner MGE transposase aligned with said sequence SEQ ID NO:
- 2, said corresponding residues being selected from the group consisting of:
- a serine;
- and
- a threonine,
- wherein said conservative substitution does not comprise substituting said phosphorylatable residue with a residue selected from the group consisting of serine, threonine, aspartic acid, glutamic acid, and proline.
Dependencies are inferred from the claim text.