A method for obtaining circular mutated and/or chimaeric polynucleotides

摘要

A quick, simple and efficient in vitro method for generating a random chimaeric and/or mutated gene or genes in a circular polynucleotide molecule is described. The method is based on hybridization of random DNA fragments to a closed, circular DNA template, elongation of the fragments by a DNA polymerase and ligation of the fragments to each other by a DNA ligase resulting in a recombined, closed circular DNA molecule which functions as a new template in a next cycle of chimaerization. The degree of recombination can be controlled by the number of cycle repetitions, the amount of different templates or/and fragments and the ratio of the latter. This procedure is referred to as polymerase chain chimaerization (PCC). The PCC method is useful for generating new improved bio-molecules.