Process for preparing variant polynucleotides

Details Process for preparing variant polynucleotides Process for preparing variant polynucleotides

2008-07-22

2008-07-22

Horlick, Kenneth R. (Department: 1637)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

C435S091200

Reexamination Certificate

active

10484682

ABSTRACT:
The present invention discloses a process for the preparation of variant polynucleotides using a combination of mutagenesis of a starting population of polynucleotides and recombination of the mutated polynucleotides. The process comprises the steps of subjecting a population of polynucleotides to (a series of) two (or more) separate PCR's comprising a first PCR with a forward mutation-specific primer for a position to be mutated and a reverse universal primer and a second PCR with a forward universal primer and a reverse mutation-specific primer for a position to be mutated. The products of the (series of) two (or more) PCR's are assembled by a polymerase, preferably in one tube.

REFERENCES:
patent: 6153410 (2000-11-01), Arnold et al.
patent: 6159690 (2000-12-01), Borrebaeck et al.
patent: 6242222 (2001-06-01), Gifford
patent: 6767701 (2004-07-01), Vind
patent: 2005/0130140 (2005-06-01), Bovenberg et al.
patent: 2005/0130141 (2005-06-01), Bovenberg et al.
patent: 1 108 783 (2001-06-01), None
patent: WO95/22625 (1995-08-01), None
patent: WO97/07205 (1997-02-01), None
patent: WO98/01581 (1998-01-01), None
patent: WO98/31837 (1998-07-01), None
patent: WO-98/41623 (1998-09-01), None
patent: WO01/29212 (2001-04-01), None
Hemsley et al. (“A simple method for site-directed mutagenesis using the polymerase chain reaction” Nucleic Acids Research. 1989. vol. 17, No. 16: pp. 6545-6551).
Zhou et al. (“Random mutagenesis of gene-sized DNA molecules by use of PCR with Taq polymerase” Nucleic Acid Research. 1991. vol. 19, No. 21: p. 6052).
Ho et al. (“Site-directed mutagenesis by overlap extension using the polymerase chain reaction” Gene. Apr. 15, 1989;77(1):51-9).
Horton et al. “Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension” Gene. Apr. 15, 1989;77(1):61-8.
Coco et al., Nature Biotechnology (2001) 19(4):354-359.
Gibbs et al., Gene, Elsevier Biomedical Press (2001) 271(1):13-20.
Kikuchi et al., Gene, Elsevier Biomedical Press (2000) 243(1-2):133-137.
Volkov et al., Nucleic Acids Research, Oxford University Press (1999) 27(18):e18.
Fromant et al., Analytical Biochemistry (1995) 224:347-353.
Ge and Rudolph, BioTechniques (1997) 22:28-30.
Higuchi et al., Nucleic Acids Research (1988) 16(15):7351-7367.
Horton et al., Gene (1989) 77:61-68.
Ho et al., Gene (1989) 77:51-59.
Landt et al., Gene (1990) 96:125-128.
Li and Shapiro, Nucleic Acids Research (1993) 21(16):3745-3748.
Picard et al., Nucleic Acids Research (1994) 22(13):2587-2591.
Sambrook and Russell, Molecular Cloning: A Laboratory Manual, Third Edition, vol. 2, Cold Spring Harbor Laboratory Press, New York (2001) pp. 13.36-13.39 and 13.79-13.80.

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