Abstract: The invention relates to a novel method for altering the sequence of a nucleic acid molecule using repair recombination in a simple one component system. The frequency of the recombination reaction is high, allowing a range of feasible selection strategies to identify successful recombination events. The method involves the steps of bringing a first nucleic acid molecule into contact with a second nucleic acid molecule in the presence of a phage annealing protein into contact with a second nucleic acid molecule in the presence of a phage annealing protein, or a functional equivalent or fragment thereof, wherein said first nucleic acid molecule comprises at least two regions of shared sequence homology with the second nucleic acid molecule, under conditions suitable for repair recombination to occur between said first and second nucleic acid molecules; ad selecting a nucleic acid molecule whose sequence has been altered so as to include sequence from said second nucleic acid molecule.

Abstract: The present invention is directed to methods and compositions for TnpI-mediated genetic engineering using the Bacillus thuringiensis recombinase TnpI and the TnpI recombination substrates TRT or TRT′, and variants thereof. In particular, the invention relates to TRT or TRT′ sequences, and variants thereof, vectors, cells and kits useful for TnpI-mediated genetic recombination, as well as methods for the use of TRT or TRT′ sequences for TnpI-mediated genetic engineering.

Abstract: The invention refers to a novel method for cloning DNA molecules using a homologous recombination mechanism between at least two DNA molecules comprising: a) providing a host cell capable of performing homologous recombination, b) contacting in said host cell a first DNA molecule which is capable of being replicated in said host cell with a second DNA molecule comprising at least two regions of sequence homology to regions on the first DNA molecule, under conditions which favour homologous recombination between said first and second DNA molecules and c) selecting a host cell in which homologous recombination between said first and second DNA molecules has occurred.

Abstract: The invention relates to methods for cloning DNA molecules using recE/recT-mediated homologous recombination mechanism between at least two DNA molecules where one DNA molecule is a circular or linear DNA molecule and the second DNA molecule is a circular DNA molecule, and the second DNA molecule contains two regions with sequence homology to the first DNA molecule. Competent cells and vectors are also described.

Abstract: The present invention is directed to methods and compositions for DNA subcloning using bacterial recombinase-mediated homologous recombination. The invention relates to methods for cloning, compositions comprising polynucleotides usefall as cloning vectors, cells comprising such polynucleotide compositions, and kits useful for cloning mediated by bacterial recombinases, such as RecE/T and Red&agr;/&bgr;.