Experimental Manipulation of Gene Expression

Contributors

Preface


1. Use of Phage λ Regulatory Signals to Obtain Efficient Expression of Genes in Escherichia Coli

I. Introduction

II. Expression of Prokaryotic Gene Products

III. Expression of Eukaryotic Genes

References


2. Multipurpose Expression Cloning Vehicles in Escherichia Coli

I. Introduction

II. pIN-I Vectors

III. pIN-II Vectors

IV. pIN-III Vectors

V. pIM Vectors: High-Copy-Number Vectors

VI. pIC Vectors: Hybrid Expression Vectors

VII. Promoter-Proving Vectors

VIII. General Cloning Strategy

IX. Summary

References


3. Molecular Cloning in Bacillus Subtilis

I. Introduction

II. Plasmid Transformation

III. Plasmid Vectors

IV. Cloning Stratagems

V. Expression of Cloned Genes

VI. Conclusions

References


4. Developments in Streptomyces Cloning

I. Introduction

II. Vectors

III. Use of Tn5 in Relation to Streptomyces DNA

IV. Applications of DNA Cloning in Streptomyces

V. Concluding Remarks

References


5. Vectors for High-Level, Inducible Expression of Cloned Genes in Yeast

I. Introduction

II. Materials and Methods

III. Results and Discussion

IV. Summary

Appendix: Plasmid Construction

References


6. Genetic Engineering of Plants by Novel Approaches

I. Introduction

II. Novel Approaches to Creating Genetic Diversity

III. Concluding Remarks

References


7. XSV2, a Plasmid Cloning Vector that Can Be Stably Integrated in Escherichia Coli

I. Introduction

II. Materials and Methods

III. Results

IV. Discussion

References


8. Construction of Highly Transmissible Mammalian Cloning Vehicles Derived from Murine Retroviruses

I. Introduction

II. General Strategy

III. Construction of a Prototype Retrovirus Vector

IV. Rescue of Recombinant Genomes as Infectious Virus

V. Characteristics of Retrovirus-Mediated Transformation

VI. Useful Derivative Vectors

VII. Conclusions and Prospects

References


9. Use of Retro virus-Derived Vectors to Introduce and Express Genes in Mammalian Cells

I. Introduction

II. Organization of the M-MuLV Genome

III. Use of Retrovirus Vectors to Study the Mechanism of Gene Expression of the M-MuLV Genome

IV. A General Transduction System Derived from the M-MuLV Genome

V. Summary and Prospects

References


10. Production of Posttranslationally Modified Proteins in the SV40-Monkey Cell System

I. Introduction

II. SV40 Late-Replacement Vectors

III. Human Growth Hormone

IV. Hepatitis B Surface Antigen

V. Conclusions and Prospects

References


11. Adenovirus Type 5 Region-EIA Transcriptional Control Sequences

I. Introduction

II. Deletion Mutations in the 5'-Flanking Sequences of Ad5 Region E1A

III. Analysis of Mutagenized Templates in Cell-Free Transcription Extracts

IV. Analysis of Cytoplasmic E1A mRNAs Found In Vivo after Infection with Deletion Mutants

V. 5'-End Analyses of E1A mRNAs Synthesized In Vivo after Infection with Deletion Mutants

VI. E1A Transcriptional Control Region and Comparison to Other Eukaryotic Control Regions

References


12. Expression of Proteins on the Cell Surface Using Mammalian Vectors

I. How Proteins Are Normally Expressed on Mammalian Cell Surfaces

II. Why It Would Be Useful to Express Proteins on the Surface of the Mammalian Cell

III. Hemagglutinin of Influenza Virus Is the Best-Characterized Integral Membrane Protein

IV. The Gene Coding for Hemagglutinin Is of Simple Structure

V. Vector Systems

VI. Hemagglutinin Is Efficiently Expressed from Both the Early and Late SV40 Promoters

VII. Small-t Intron Leads to Genetic Instability of the Early-Replacement, Vector

VIII. Hemagglutinin Synthesized by SV40-HA Recombinants is Biologically Active

IX. Removing the C-Terminal Hydrophobie Sequence Converts Hemagglutinin from an Integral Membrane Protein to a Secreted Protein

X. Prospects

References


13. Expression of Human Interferon-7 in Heterologous Systems

I. Introduction

II. Structure of the Human Interferon-7 cDNA

III. Heterologous Expression in Escherichia Coli

IV. Expression in the Yeast Saccharomyces Cerevisiae

V. Conclusion

References


14. Commercial Production of Recombinant DNA-Derived Products

I. Introduction

II. Production of Biosynthetic Human Insulin

III. Other Pharmaceutical Applications of Recombinant DNA

IV. Conclusion

References

Appendix 1. Two-Dimensional DNA Electrophoretic Methods Utilizing in Situ Enzymatic Digestions

I. Introduction

II. Experimental Procedures

III. Examples

IV. Conclusion

References

Appendix 2. Site-Specific Mutagenesis Using Synthetic Oligodeoxyribonucleotides as Mutagens

I. Introduction

II. Experimental Procedures

III. Example

IV. Conclusion

References

Index