Article
DNA shuffling method for generating highly recombined genes and evolved enzymes.
Nature biotechnology - 1 Apr 2001
Coco W M, Levinson W E, Crist M J, Hektor H J, Darzins A, Pienkos P T, Squires C H, Monticello D J
Abstract excerpt
We introduce a method of in vitro recombination or "DNA shuffling" to generate libraries of evolved enzymes. The approach relies on the ordering, trimming, and joining of randomly cleaved parental DNA fragments annealed to a transient polynucleotide scaffold. We generated chimeric libraries averaging 14.0 crossovers per gene, a several-fold higher level of recombination than observed for other methods. We also...
Topics
- Alleles
- Amino Acid Sequence
- Crossing Over, Genetic
- DNA, Complementary
- Gene Library
- Genetic Techniques
- Molecular Sequence Data
- Mutagenesis
- Mutation
- Nocardia
- Polymerase Chain Reaction
- Polymorphism, Restriction Fragment Length
- Recombination, Genetic
- Rhodococcus
- Sequence Homology, Amino Acid
- Substrate Specificity