Gene Expression Systems: Using Nature for the Art of Expression
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Table of contents
- Gene Expression Systems: Using Nature for the Art of Expressioniii
- Copyright Pageiv
- Contentsv
- Contributorsxi
- Introduction1
- The Art of Expression1
- Purpose of This Book2
- Selecting a Suitable Expression System: Considerations3
- Genomics and the Future of Protein Expression Systems5
- SECTION I: PROKARYOTIC EXPRESSION SYSTEMS7
- Chapter 1. Gene Expression Systems Based on Bacteriophage T7 RNA Polymerase9
- Introduction10
- T7 RNAP in Its Natural Habitat11
- Biochemistry of T7 RNAP14
- T7-Based Gene Expression Systems24
- Conclusions36
- References37
- Chapter 2. Expression Vectors Employing the trc Promoter45
- Introduction46
- Vectors for trc-Driven Expression51
- Additional Hosts for trc-Driven Expression57
- Outlook for Further Improvements of Expression Vectors Employing the trc Promoter58
- References59
- Chapter 3. Bacillus Expression" A Gram-Positive Model65
- Introduction66
- Bacilli Used as Industrial Production Organisms67
- Genetic Manipulation in Bacilli71
- Plasmid Vectors75
- Transcriptional Regulation77
- Conclusions81
- Appendix82
- References89
- Chapter 4. araB Expression System in Escherichia coli95
- Introduction95
- The araB Promoter and How It Works97
- Use of the ara System for Expression of Recombinant Products99
- Conclusions104
- References106
- SECTION II: EUKARYOTIC EXPRESSION SYSTEMS109
- Chapter 5. Adenoviral Vectors for Protein Expression111
- Adenovirus Biology112
- Adaptation of Adenovirus as a Gene Transfer Vector117
- Construction of Adenovirus Expression Vectors119
- Applications of Adenoviral Vectors in Gene Expression128
- Considerations in the Use of Adenovirus Expression Vectors138
- Advantages and Disadvantages of Adenoviral Vectors141
- Future Directions143
- Conclusions144
- References145
- Chapter 6. Expression in the Methylotrophic Yeast Pichia pastoris157
- Introduction158
- Background Information161
- Construction of Expression Strains166
- Important Considerations in the Use of the Expression System175
- Conclusion183
- References184
- Chapter 7. Recombinant Protein Expression in Pichia methanolica193
- Background193
- DNA Transformation194
- Expression Vector and Identification of an Expression Strain196
- Gene Disruptions and Generation of a Protease-Deficient Strain198
- Fermentation199
- Evaluation of Protein Expression199
- Expression of the 65-kDa Isoform of Human Glutamate202
- Protein Secretion202
- Benefits and Liabilities of the P. methanolica Expression System205
- Summary206
- Appendix207
- References207
- Chapter 8. Cytomegalovirus Promoter for Expression in Mammalian Cells211
- Introduction212
- The Cytomegalovirus Enhancer-Containing Promoter214
- Positive Regulation216
- Experimental Procedures219
- Effects Upstream of the Cytomegalovirus Enhancer221
- Effects Downstream of the Cytomegalovirus Promoter222
- Negative Regulation223
- Conclusions225
- References228
- Chapter 9. Inducible Mammalian Expression Systems235
- Background: Development of Inducible Expression Systems235
- Literature Review245
- Author's Experience248
- Comparing Inducible Mammalian Expression Systems Currently Available250
- Conclusions and Future Directions250
- References253
- Chapter 10. Protein Expression in Mammalian Cells Using Sindbis Virus259
- Introduction260
- Applications267
- Protein Expression272
- Comparisons with Other Systems279
- New Directions and Conclusions282
- References283
- SECTION III: EXPRESSION IN INSECT SYSTEMS287
- Chapter 11. Drosophila S2 System for Heterologous Gene Expression289
- Introduction290
- Properties of S2 Line290
- Experimental Procedures292
- Literature Review300
- Choosing an Expression System315
- Future Directions318
- Conclusions321
- Appendix321
- References325
- Chapter 12. Baculovirus Expression Vector System331
- Introduction332
- Background334
- Basic Research of Baculoviruses as a Tool for Gene Expression347
- Expression Examples349
- Guidelines to Optimize Heterologous Gene Expression in Baculovirus352
- Summary and Future354
- Appendix355
- References359
- SECTION IV: TRANSGENIC EXPRESSION365
- Chapter 13. Recombinant Protein Expression in Transgenic Mice367
- Overview and Background368
- Transgenic Mice: General Overview368
- Examples of Recombinant Protein Expression in Transgenic Mice371
- Modeling Human Disease through Targeted Overexpression378
- Biomass: Transgenic Mice as Model Bioreactors381
- Future of Transgenic Mice Approaches: Need for Inducible Expression383
- Conclusions390
- References391
- Chapter 14. Expression of Recombinant Proteins in the Milk of Transgenic Animals399
- Introduction400
- Expression of Heterologous Proteins in Milk401
- Milk-Specific Transgenes403
- Insertion of the Transgene into the Germ Line405
- Transgenic Animal Production408
- Biosynthesis of Milk Proteins409
- Milk Secretion from the Mammary Gland412
- Lactation and Milk Output412
- Milk Composition and Purification of the Target Protein413
- Quality Issues in Transgenic Production417
- Regulatory Considerations418
- Current Status and Future Directions420
- References421
- Chapter 15. Recombinant Protein Expression in Plants429
- General Introduction430
- Transformation Methods431
- Promoters Used for Recombinant Protein Accumulation in Plants435
- Expression of Recombinant Proteins in Plants and Agricultural Biotechnology438
- Recombinant Protein Expression in Plants to Obtain New Products442
- Protein Targeting and Accumulation447
- Virus-Mediated Expression Systems448
- Summary/Discussion450
- References452
- Index463
Gene Expression Systems: Using Nature for the Art of Expression offers detailed information on a wide variety of gene expression systems from an array of organisms. It describes several different types of expression systems including transient, stable, viral, and transgenic systems. Each chapter is written by a leader in the field. The book includes timelines and examples for each expression system, and provides an overview of the future of recombinant protein expression.
Key Features
* Provides detailed information on expression systems
* Covers a variety of promoters and host organisms enabling researchers to tailor protocols to their specific needs
* Includes timelines and examples
* Compares pros and cons of each method
Key Features
* Provides detailed information on expression systems
* Covers a variety of promoters and host organisms enabling researchers to tailor protocols to their specific needs
* Includes timelines and examples
* Compares pros and cons of each method