Journal
IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-PLANT
Volume 36, Issue 5, Pages 331-337Publisher
SOC IN VITRO BIOLOGY
DOI: 10.1007/s11627-000-0060-0
Keywords
DNA shuffling; exon shuffling; molecular breeding; sexual PCR
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DNA shuffling is a technique being utilized for in vitro recombination of a single gene or pools of homologous genes. The genes are fragmented into randomly sized pieces, and polymerase chain reaction (PCR) reassembly of full-length genes front the fragments, via self-priming, yields recombination due to PCR template switching. After these PCR products are screened and the interesting products sequenced, improved clones are reshuffled to recombine useful mutations in additive or synergistic ways, in effect mimicking the process of natural sexual recombination. Protein can be 'bred' with the appropriate individual properties and then their 'progeny' screened for the desired combination of traits. DNA shuffling is a powerful tool enabling rapid and directed evolution of new genes, operons and whole viral genomes.
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