| 摘要 |
A method for replicating and amplifying a target nucleic acid sequence is described. A method of the invention involves the formation of a recombinati on intermediate without the prior denaturing of a nucleic acid duplex through t he use of a recombination factor. The recombination intermediate is treated wit h a high fidelity polymerase to permit the replication and amplification of th e target nucleic acid sequence. In preferred embodiments, the polymerase comprises a polymerase holoenzyme. In further preferred embodiments, the recombination factor is bacteriophage T4 UvsX protein or homologs from other species, and the polymerase holoenzyme comprises a polymerase enzyme, a clam p protein and a clamp loader protein, derived from viral, bacteriophage, prokaryotic, archaebacterial, or eukaryotic systems. |
