Summary: © 1998 Oxford University Press 681­683Nucleic Acids Research, 1998, Vol. 26, No. 2
Random-priming in vitro recombination: an effective
tool for directed evolution
Zhixin Shao, Huimin Zhao, Lori Giver and Frances H. Arnold*
Division of Chemistry and Chemical Engineering 210-41, California Institute of Technology, Pasadena,
CA 91125, USA
Received September 15, 1997; Revised and Accepted November 21, 1997
ABSTRACT
A simple and efficient method for in vitro mutagenesis
and recombination of polynucleotide sequences is
reported. The method involves priming template
polynucleotide(s) with random-sequence primers and
extending to generate a pool of short DNA fragments
which contain a controllable level of point mutations.
The fragments are reassembled during cycles of
denaturation, annealing and further enzyme-catalyzed
DNA polymerization to produce a library of full-length
sequences. Screening or selecting the expressed gene
products leads to new variants with improved
functions, as demonstrated by the recombination of

Source: Arnold, Frances H. - Division of Chemistry and Chemical Engineering, California Institute of Technology

Collections: Chemistry; Biology and Medicine