HyperMu™ MuA Transposase

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Applications

• In vitro insertion of an artificial Mu transposon into DNA cloned in vectors, such as plasmids, fosmids, cosmids, or BACs, as well as in vitro insertion into linear DNA.
• Preparation of HyperMu Transposomes for in vivo transposition* of an artifical Mu transposon following electroporation into living cells.

HyperMu™ MuA Transposase is a hyperactive enzyme that is at least 50 times more active than native MuA transposase available from other suppliers of Mu-based systems. The highly purified, single-subunit HyperMu MuA Transposase can be used to randomly insert (transpose or "hop") any HyperMu™ Transposon into any target DNA in vitro with an efficiency of up to >10⁶ insertion clones per standard reaction. HyperMu Transposase can also catalyze transposition of other artificial Mu transposons that have the R1 and R2 recognition sequences in the proper orientation at each end. The DNA between the R1 and R2 transposon end sequences can be any DNA, including DNA encoding selectable markers, origins of replication, or other genetic elements.

Further, when HyperMu MuA Transposase is incubated with an artificial Mu transposon in the absence of Mg²⁺, a stable HyperMu Transposome complex is formed. Like an EZ-Tn5™ Transposome™ complex, a HyperMu Transposome can be electroporated into living cells, where it is activated by intracellular Mg²⁺ and the Mu transposon component is randomly inserted into the host's DNA.

A typical HyperMu transposition reaction requires four components: 1) the HyperMu Transposase; 2) an artificial Mu transposon; 3) a target DNA; and 4) the presence of Mg²⁺. Insertion of an artificial Mu transposon into the target DNA results in a 5-bp duplication of target DNA sequence immediately adjacent to both ends of the transposon.

Benefits

• At least 50 times more active than the MuA transposase available from other suppliers.
• HyperMu MuA Transposase recognizes the same R1 and R2 transposon recognition sequences as MuA transposase and is therefore compatible with other artificial Mu transposons.

*The use of Transposome™ complexes for in vivo insertion of a transposon, including, but not limited, to HyperMu™ and EZ-Tn5™ Transposome™ complexes, is covered by U.S. Patent No. 6,159,736 and related patents and patent applications, exclusively licensed to EPICENTRE.