PMID: 9421535Feb 28, 1998Paper

Random-priming in vitro recombination: an effective tool for directed evolution

Nucleic Acids Research
Z ShaoF H Arnold

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 genes encoding different thermostable subtilisins in order to obtain enzymes more stable than either parent.

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Citations

Oct 10, 2001·Angewandte Chemie·Sean V. TaylorDonald Hilvert
Jul 28, 2012·Biodegradation·Smriti RayuBrajesh K Singh
Dec 7, 2007·Applied Biochemistry and Biotechnology·S SenB Mandal
Mar 31, 2004·Bioorganic & Medicinal Chemistry·Eduardo García-JuncedaAlfonso Fernández-Mayoralas
Sep 11, 2002·Journal of Molecular Biology·Paul E O'MailleMing-Daw Tsai
Jun 26, 2003·Journal of Molecular Biology·Kaori Hiraga, Frances H Arnold
Jan 11, 2001·Biochimica Et Biophysica Acta·P N Bryan
Jan 11, 2001·Biochimica Et Biophysica Acta·P P SheridanJ E Brenchley
Jun 14, 2000·Current Opinion in Biotechnology·K Furukawa
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Aug 6, 1998·Current Opinion in Chemical Biology·L Giver, F H Arnold
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Feb 18, 1999·Current Opinion in Chemical Biology·F H Arnold, A A Volkov
May 1, 2001·Nature Biotechnology·V SieberF H Arnold
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Apr 29, 2005·Biomolecular Engineering·Linda G Otten, Wim J Quax

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