Enzymatic inverse polymerase chain reaction library mutagenesis

Details Enzymatic inverse polymerase chain reaction library mutagenesis Enzymatic inverse polymerase chain reaction library mutagenesis

1994-06-01

1996-04-30

Jones, W. Gary

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

435 691, 435 6, 536 241, 536 2433, 935 77, 935 78, C12P 1934, C12P 2100, C07H 2104, C12Q 168

Patent

active

055124631

ABSTRACT:
This invention discloses a method for generating a recombinant library by introducing one or more changes within a predetermined region of double-stranded nucleic acid, comprising providing a first primer population and a second primer population, each of the populations having a variable base composition at known positions along the primers, the primers incorporating a class IIS restriction enzyme recognition sequence, being capable of directing change in the nucleic acid sequence and being substantially complementary to the double stranded nucleic acid to permit hybridization thereto. The method additionally comprises hybridizing the first and second primer populations to opposite strands of the double stranded nucleic acid to form a first pair of primer-templates oriented in opposite directions, performing enzymatic inverse polymerase chain reaction to generate at least one linear copy of the double stranded nucleic acid incorporating the change directed by the primers, cutting the double stranded nucleic acid copy with a class IIS restriction enzyme to form a restricted linear nucleic acid molecule containing the change, joining termini of the restricted linear nucleic acid molecule to produce double-stranded circular nucleic acid and introducing the nucleic acid into compatible host cells. A method is additionally provided for generating a recombinant library using wobble-base mutagenesis.

REFERENCES:
patent: 4800159 (1989-01-01), Mullis et al.
patent: 4935357 (1990-06-01), Szybalski
patent: 4959312 (1990-09-01), Sirotkin
patent: 5023171 (1991-06-01), Ho et al.
Hermes, et al. "Searching Sequence Space by Definably Random Mutagenesis: Improving the Catalytic Potency of an Enzyme": Proc. Natl. Acad. Sci. 87: 696-700 (1990).
Higuchi, et al. "A General Method of in vitro Preparation and Specific Mutagenesis of DNA Fragment: Study of Protein and DNA Interactions" Nucl. Acids Res. 16(15): 7351-7367 (1988).
Jones, et al. "A Rapid Method for Site-Specific Mutagenesis and Directional Subcloning by Using the Polymerase Chain Reaction to Generate Recombinant Circles" BioTechniques 8: 178-183 (1990).
Jones, et al. "A Rapid Method for Recombination and Site-Specific Mutagenesis by Placing Homologous Ends in DNA Using Polymerase Chain Reaction" BioTechniques 10: 62-66 (1991).
Kozak, M. "Compilation and Analysis of Sequences Upstream from the Translational Start Site in Eukaryotic mRNAs": Nucl. Acids Res. 12(2): 857-872 (1984).
Movva, et al. "Amino Acid Sequence of the Signal Peptide of ompA Protein, a Major Outer Membrane Protein of Escherichia coli" J. of Biol. Chem. 255: 27-29 (1980).
Movva, et al. "Gene Structure of the OmpA Protein, a Major Surface Protein of Escherichia coli Required for Cell--Cell Interaction" J. Mol. Biol. 143: 317-328 (1980).
Nelson, et al. "A General Method of Site-Specific Mutagenesis Using a Modification of the Thermus aquaticus Polymerase Chain Reaction" Anal. Biochem. 180: 147-151 (1989).
Saiki, et al. "Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA Polymerase" Science 239: 487-491 (1988).
Seed, B. "Purification of Genomic Sequence from Bacteriophase Libraries by Recombination and Selection in vivo" Nucl. Acids Res. 11(8): 2427-2455 (1983).
Skerra, et al. "Filter Screening of Antibody Fab Fragments Secreted from Individual Bacterial Colonies: Specific Detection of Antigen Binding with a Two-Membrane System" Anal. Biochem. 196:151-155 (1991).
Stemmer and Morris, "Enzymatic Inverse PCR: A Restriction Site Independent Single-Fragment Method for High-Efficiency, Site-Directed Mutagenesis" BioTechniques 13(2): 216-200 (1992).
Jones, et al., "DNA Mutagenesis and Recombination" Nature 344: 793-794 (Apr. 1990).
Huang, et al., Vectors: A survey of Molecular Cloning Vectors and teir uses (1987) R. L. Rodriquez and D. T. Denhardt (eds.) Buttenworth Publ. pp. 269-283.
Szardenings et al. (1990) Gene, vol. 94, pp. 1-7.
Hermes, J. D. et al. (1989) Gene, vol. 84, pp. 143-151.
Tomic et al. (1990) Nucleic Acids Research, vol. 18, No. 6, pp. 1656.
Hemsley, A. et al. (1989) Nucleic Acids Research, vol. 17, No. 16, pp. 6545-6551.
Bucheler et al. (1990) Gene, vol. 96, pp. 271-276.

Affiliated with

Also associated with

Rating

Enzymatic inverse polymerase chain reaction library mutagenesis does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Enzymatic inverse polymerase chain reaction library mutagenesis, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Enzymatic inverse polymerase chain reaction library mutagenesis will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-627907