Patent ID: 7582423
Filing Date: 2009-09-01
Classification: C12N

Abstract:
1. An in vitro method of redistributing sequence variations between non-identical polynucleotides wherein each step is performed outside of living cells, comprising: a) making a heteroduplex polynucleotide from two non-identical polynucleotides having 96% or more sequence identity, the heteroduplex having first and second strands; b) mixing the heteroduplex polynucleotide with an effective amount of mismatch-directed strand cleavage activity, proofreading activity, and ligase activity wherein said mismatch-directed strand cleavage activity consists of a single enzyme that recognizes a mismatch directly; and c) allowing sufficient time for at least two non-complementary nucleotide base pairs to be converted to complementary base pairs wherein a population of polynucleotide strands not identical to either of said first and second strands results and wherein the polynucleotide strands of the population have an average of 1 or more crossovers per 120 basepairs relative to the distance between the first and last mismatches.