High throughput protein expression and purification : methods and protocols / edited by Sharon A. Doyle. — Totowa, N.J. : Humana Press, c2009. – (58.17/M592/v.498)

Contents

    Contents
    
    Preface
    Contributors
    1 High-Throughput Protein Production (HTPP): A Review of Enabling Technologies to Expedite Protein Production
    2 Designing Experiments for High-Throughput Protein Expression
    3 Gateway Cloning for Protein Expression
    4 Flexi Vector Cloning
    5 The Precise Engineering of Expression Vectors Using High-Throughput In-Fusion PCR Cloning 75
    6 The Polymerase Incomplete Primer Extension (PIPE) Method Applied to High-Throughput Cloning and Site-Directed Mutagenesis 91
    7 A Family of LIC Vectors for High-Throughput Cloning and Purification of Proteins
    8 "System 48" High-Throughput Cloning and Protein Expression Analysis 117
    9 Automated 96-Well Purification of Hexahistidine-Tagged Recombinant Proteins on MagneHis Ni2+-Particles 129
    10 E. coli and Insect Cell Expression， Automated Purification and Quantitative Analysis
    11 Hexahistidine-Tagged Maltose-Binding Protein as a Fusion Partner for the Production of Soluble Recombinant Proteins in Escherichia coli 157
    12 PHB-Intein-Mediated Protein Purification Strategy
    13 High-Throughput Biotinylation of Proteins
    14 High-Throughput Insect Cell Protein Expression Applications
    15 High-Throughput Protein Expression Using Cell-Free System 229
    16 The Production of Glycoproteins by Transient Expression in Mammalian Cells 245
    17 High-Throughput Expression and Detergent Screening of Integral Membrane Proteins
    18 Cell-Free Expression for Nanolipoprotein Particles: Building a High-Throughput Membrane Protein Solubility Platform 273
    19 Expression and Purification of Soluble His6-Tagged TEV Protease 297
    20 High-Throughput Protein Concentration and Buffer Exchange: Comparison of Ultrafiltration and Ammonium Sulfate Precipitation 309
    Index