Recombinant DNA methods are powerful, revolutionary techniques that allow the isolation of single genes in large amounts from a pool of thousands or millions of genes and the modification of these isolated genes or their regulatory regions for reintroduction into cells for expression at the RNA or protein levels. These attributes lead to the solution of complex biological problems and the production of new and better products in the areas of medicine, agriculture, and industry.
Recombinant DNA Methodology, a volume in the Selected Methods in Enzymology series produced in benchtop format, contains a selection of key articles from Volumes 68, 100, 101, 153, 154, and 155 of Methods in Enzymology. The essential and widely used procedures provided at an affordable price will be an invaluable aid to the graduate student and the researcher.
• Enzymes in DNA research
• DNA isolation, hybridization, and cloning
• DNA sequence analysis
• cDNA cloning
• Gene products
• Identification of cloned genes and mapping of genes
• Monitoring cloned gene expression
• Cloning and transferring of genes into yeast cells
• Cloning and transferring of genes into plant cells
• Cloning and transferring of genes into animal cells
• Site-directed mutagenesis
• Protein engineering
• Expression vectors
Table of Contents
Enzymes in Recombinant DNA Research:
N.P. Higgins and N.R. Cozzarelli, DNA-Joining Enzymes: A Review.
R. Fuchs and R. Blakesley, Guide to the Use of Type II Restriction Endonucleases.
M. McClelland, Site-Specific Cleavage of DNA at 8-, 9- and 10bp Sequences.
L.-H. Guo and R. Wu, Exonuclease III: Use for DNA Sequence Analysis and in Specific Deletions of Nucleotides.
Methods for Isolation, Purification, or Amplification of DNA:
E. Southern, Gel Electrophoresis of Restriction Fragments.
C.L. Smith and C.R. Cantor, Purification, Specific Fragmentation, and Separation of Large DNA Molecules.
G.F. Carle and M.V. Olson, Orthogonal-Field-Alternation Gel Electrophoresis.
H.C. Birnboim, A Rapid Alkaline Extraction Method for the Isolation of Plasmid DNA.
K.B. Mullis and F.A. Faloona, Specific Synthesis of DNA in Vitro via a Polymerase-Catalyzed Chain Reaction.
Vectors or Methods for Gene Cloning:
J. Collins, Escherichia coli Plasmids Packageable in Vitro in a γl Bacteriophage Particles.
J. Vieira and J. Messing, Production of Single-Stranded Plasmid DNA.
H. Okayama, M. Kawaichi, M. Brownstein, F. Lee, T. Yokota, and K. Arai, High-Efficiency Cloning of Full-Length cDNA; Construction and Screening of cDNA Expression Libraries for Mammalian Cells.
D.A. Morrison, Transformation and Preservation of Competent Bacterial Cells by Freezing.
D. Hanahan and M. Meselson, Plasmid Screening at High Colony Density.
J. Karn, S. Brenner, and L. Barnett, New Bacteriophage Lambda Vectors with Positive Selection for Cloned Inserts.
A.M. Frischauf, N. Murray, and H. Lehrach, γl Phage Vectors-EMBL Series.
M. Snyder, S. Elledge, D. Sweetser, R.A. Young, and R.W. Davis, γlgt 11: Gene Isolation with Antibody Probes and Other Applications:
R.J. Rothstein, One-Step Gene Disruption in Yeast.
S.W. Rudy, J.W. Szostak, and A.W. Murrayv, Cloning Regulated Yeast Genes from a Pool of lacZ Fusions.
L. Clarke, C.-L. Hsiao, and J. Carbon, Selection Procedure for Isolation of Centromere DNAs from Saccharomyces cerevisiae.
J.R. Broach, Construction of High Copy Yeast Vectors Using 2-~gmm Circle Sequences.
S.G. Rogers, H.J. Klee, R.B. Horsch, and R.T. Fraley, Improved Vectors for Plant Transformation: Expression Cassette Vectors and New Selectable Markers.
Vectors or Methods for Expression of Cloned Genes:
M.J. Casadaban, A. Martinez-Arias, S.K. Shapira, and J. Chou, β-Galactosidase Gene Fusions for Analyzing Gene Expression in Escherichia coli and Yeast.
M. Rosenberg, Y.-S. Ho, and A. Shatzman, The Use of pKC30 and Its Derivatives for Controlled Expression of Genes.
E. Remaut, A. Marmenout, G. Simons, and W. Fiers, Expression of Heterologous Unfused Protein in Escherichia coli.
G.D. Duffaud, P.E. March, and M. Inouye, Expression and Secretion of Foreign Proteins in Escherichia coli.
L. Guarente, Yeast Promoters and LacZ Fusions Designed to Study Expression of Cloned Genes in Yeast.
J.B. Gurdon and M.P. Wickens, The Use of Xenopus Oocytes for the Expression of Cloned Genes.
G.A. Bitter, K.M. Egan, R.A. Koski, M.O. Jones, S.G. Elliott, and J.C. Giffin, Expression and Secretion Vectors for Yeast.
Methods for Oligonucleotide-Directed Mutagenesis:
M.J. Zoller and M. Smith, Oligonucleotide-Directed Mutagenesis of DNA Fragments Cloned into M13 Vectors.
W. Kramer and H.-J. Fritz, Oligonucleotide-Directed Construction of Mutations via Gapped Duplex DNA.
T.A. Kunkel, J.D. Roberts and R.A. Zakour, Rapid and Efficient Site-Specific Mutagenesis without Phenotypic Selection.
R. Pine and P.C. Huang, An Improved Method to Obtain a Large Number of Mutants in a Defined Region of DNA.
D.E. Hill, A.R. Oliphant, and K. Struhl, Mutagenesis with Degenerate Oligonucleotides: An Efficient Method for Saturating a Defined DNA Region with Base Pair Substitutions.
A. Vainstein, A. Razin, A. Graessmann, and A. Loyter, Fusogenic Reconstituted Sendai Virus Envelopes as a Vehicle for Introducing DNA into Viable Mammalian Cells.
J.L. Manley, A. Fire, M. Samuels, and P.A. Sharp, In Vitro Transcriptions: Whole-Cell Extract.
W.C. Merrick, Translation of Exogenous mRNAs in Reticulocyte Lysates.
C.W. Anderson, J.W. Straus, and B.S. Dudock, Preparation of a Cell-Free Protein-Synthesizing System from Wheat Germ.
R.D. Shillito and I. Potrykus, Direct Gene Transfer to Protoplasts of Dicotyledonous and Monocotyledonous Plants by a Number of Methods, Including Electroporation.
R.M.K. Dale and A. Arrow, A Rapid Single-Stranded Cloning, Sequencing, Insertion, and Deletion Strategy.
T.D. Tullius, B.A. Dombroski, M.E.A. Churchhill, and L. Kam, Hydroxyl Radical Footprinting: A High-Resolution Method for Mapping Protein-DNA Contacts.