E. coli Gene Expression Protocols

1. Front Matter

   Pages i-xi

   PDF

2. Cold-Inducible Promoters for Heterologous Protein Expression

   • François Baneyx, Mirna Mujacic

   Pages 1-18

3. Dual-Expression Vectors for Efficient Protein Expression in Both E. coli and Mammalian Cells

   • Rebecca L. Mullinax, David T. Wong, Heidi A. Davis, Kerstein A. Padgett, Joseph A. Sorge

   Pages 19-30

4. A Dual-Expression Vector Allowing Expression in E. coli and P. pastoris, Including New Modifications

   • Angelika Lueking, Sabine Horn, Hans Lehrach, Dolores J. Cahill

   Pages 31-42

5. Purification of Recombinant Proteins from E. coli by Engineered Inteins

   • Ming-Qun Xu, Thomas C. Evans Jr.

   Pages 43-68

6. Calmodulin as an Affinity Purification Tag

   • Samu Melkko, Dario Neri

   Pages 69-77

7. Calmodulin-Binding Peptide as a Removable Affinity Tag for Protein Purification

   • Wolfgang Klein

   Pages 79-97

8. Maltose-Binding Protein as a Solubility Enhancer

   • Jeffrey D. Fox, David S. Waugh

   Pages 99-117

9. Thioredoxin and Related Proteins as Multifunctional Fusion Tags for Soluble Expression in E. coli

   • Edward R. LaVallie, Elizabeth A. DiBlasio-Smith, Lisa A. Collins-Racie, Zhijian Lu, John M. McCoy

   Pages 119-140

10. Discovery of New Fusion Protein Systems Designed to Enhance Solubility in E. coli

    • Gregory D. Davis, Roger G. Harrison

    Pages 141-154

11. Assessment of Protein Folding/Solubility in Live Cells

    • Rhesa D. Stidham, W. Christian Wigley, John F. Hunt, Philip J. Thomas

    Pages 155-170

12. Improving Heterologous Protein Folding via Molecular Chaperone and Foldase Co-Expression

    • François Baneyx, Joanne L. Palumbo

    Pages 171-197

13. High-Throughput Purification of PolyHis-Tagged Recombinant Fusion Proteins

    • Thomas Lanio, Albert Jeltsch, Alfred Pingoud

    Pages 199-204

14. Co-Expression of Proteins in E. coli Using Dual Expression Vectors

    • Karen Johnston, Ronen Marmorstein

    Pages 205-213

15. Small-Molecule Affinity-Based Matrices for Rapid Protein Purification

    • Karin A. Hughes, Jean P. Wiley

    Pages 215-223

16. Use of tRNA-Supplemented Host Strains for Expression of Heterologous Genes in E. coli

    • Carsten-Peter Carstens

    Pages 225-233

17. Screening Peptide/Protein Libraries Fused to the λ Repressor DNA-Binding Domain in E. coli Cells

    • Leonardo Mariño-RamÍrez, Lisa Campbell, James C. Hu

    Pages 235-250

18. Studying Protein-Protein Interactions Using a Bacterial Two-Hybrid System

    • Simon L. Dove

    Pages 251-265

19. Using Bio-Panning of FLITRX Peptide Libraries Displayed on E. coli Cell Surface to Study Protein-Protein Interactions

    • Zhijian Lu, Edward R. LaVallie, John M. McCoy

    Pages 267-280

20. Use of Inteins for the In Vivo Production of Stable Cyclic Peptide Libraries in E. coli

    • Ernesto Abel-Santos, Charles P. Scott, Stephen J. Benkovic

    Pages 281-294

Peter E. Vaillancourt presents a collection of popular and emerging methodologies that take advantage of E. coli's ability to quickly and inexpensively express recombinant proteins. The authors focus on two areas of interest: the use of E. coli vectors and strains for production of pure, functional protein, and the use of E. coli as host for the functional screening of large collections of proteins and peptides. Among the cutting-edge techniques demonstrated are those for rapid high-level expression and purification of soluble and functional recombinant protein and those essential to functional genomics, proteomics, and protein engineering.

• Promoter
• Proteomics
• biosynthesis
• gene expression
• genes
• tRNA
• transcription

"...many will find it extremely useful to have this book on hand when trying to achieve high levels of pure, soluble recombinant protein; or incorporating an E. coli expression step into the procedures of functional genomics, proteomics and protein engineering." - Microbiology Today

• Applied Molecular Evolution, San Diego, USA

  Peter E. Vaillancourt

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