title: DNA shuffling of cytochrome P450 enzymes.
abstract: DNA family shuffling is an efficient method for creating libraries of novel enzymes, in which a high proportion of mutants exhibit correct folding and possess catalytic properties distinct from the starting material. The evolutionary arrangement of cytochromes P450 into subfamilies of enzymes with highly similar nucleotide sequences but distinct catalytic properties renders them excellent starting material for DNA family shuffling experiments. This chapter provides a general method for creating libraries of shuffled P450s from two or more related sequences and incorporates several recent improvements to previously published methods.
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, Australia.
authors.forename: James B Y H
authors.initials: JB
authors.lastname: Behrendorff
authors.forename: Wayne A
authors.initials: WA
authors.lastname: Johnston
authors.forename: Elizabeth M J
authors.initials: EM
authors.lastname: Gillam
mesh_terms: D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D032621:DNA Shuffling / Q000379:methods*; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction
chemical_list: D018076:DNA, Complementary; D003577:Cytochrome P-450 Enzyme System