RANDOM-PRIMING IN-VITRO RECOMBINATION - AN EFFECTIVE TOOL FOR DIRECTED EVOLUTION

Citation
Zx. Shao et al., RANDOM-PRIMING IN-VITRO RECOMBINATION - AN EFFECTIVE TOOL FOR DIRECTED EVOLUTION, Nucleic acids research, 26(2), 1998, pp. 681-683
Citations number
13
Categorie Soggetti
Biology
Journal title
ISSN journal
03051048
Volume
26
Issue
2
Year of publication
1998
Pages
681 - 683
Database
ISI
SICI code
0305-1048(1998)26:2<681:RIR-AE>2.0.ZU;2-E
Abstract
A simple and efficient method for in vitro mutagenesis and recombinati on of polynucleotide sequences is reported. The method involves primin g template polynucleotide(s) with random-sequence primers and extendin g to generate a pool of short DNA fragments which contain a controllab le level of point mutations. The fragments are reassembled during cycl es of denaturation, annealing and further enzyme-catalyzed DNA polymer ization to produce a library of full-length sequences. Screening or se lecting the expressed gene products leads to new variants with improve d functions, as demonstrated by the recombination of genes encoding di fferent thermostable subtilisins in order to obtain enzymes more stabl e than either parent.