| Description |
1 online resource |
| Series |
Learning materials in biosciences |
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Learning materials in biosciences.
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| Contents |
Intro -- Preface -- Motivation -- Target Audience and Prerequisites -- Writing Styles -- The Case Study -- The Organization -- The Companion Website -- Feedback -- Acknowledgement -- Contents -- About the Authors -- 1: Literature Search -- 1.1 Databases -- 1.2 How to Search? -- 1.3 Important Information -- Exercise -- Further Reading -- 2: Sequence Analysis -- 2.1 Gene Information -- 2.1.1 Kyoto Encyclopedia of Genes and Genomes (KEGG) -- 2.1.2 National Center for Biotechnology Information (NCBI) -- 2.1.3 SnapGene -- 2.1.4 Other Databases of Protein Sequences -- 2.2 Gene Analysis |
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2.2.1 Protein Sequence Analysis -- 2.2.1.1 ProtParam -- 2.2.1.2 SignalP -- 2.2.1.3 BLAST -- 2.2.1.4 Clustal Omega -- 2.2.1.5 Pfam -- 2.2.1.6 PROSITE -- 2.2.1.7 SMART -- 2.2.1.8 InterPro -- 2.2.2 DNA Sequence Analysis -- 2.2.2.1 Reverse Complement -- 2.2.2.2 DNA Stats -- 2.2.2.3 Translate -- 2.2.2.4 Codon Usage -- 2.2.2.5 Reverse Translate -- 2.2.3 Perl -- Exercise -- Further Reading -- 3: Structural Analysis -- 3.1 Protein Structure Databases -- 3.1.1 Protein Data Bank (PDB) -- 3.1.2 Biological Magnetic Resonance Data Bank (BMRB) -- 3.2 Protein Structure Prediction |
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3.2.1 Protein Secondary Structure Prediction -- 3.2.1.1 JPred -- 3.2.1.2 PSIPRED -- 3.2.1.3 PredictProtein -- 3.2.2 Protein Tertiary Structure Prediction -- 3.2.2.1 Template-Based Modelling -- 3.2.2.2 Template-Free Modelling -- 3.3 Protein Structure Visualization Tools -- Exercise -- Further Reading -- 4: Protein Expression Hosts and Expression Plasmids -- 4.1 Escherichia coli Strains -- 4.2 Plasmid and Plasmid Map -- 4.2.1 Finding a Plasmid -- 4.2.2 Reading a Plasmid Map -- 4.2.3 Creating a Plasmid Map -- 4.2.4 Plasmid Selection -- 4.2.4.1 Promoter -- 4.2.4.2 Protein Tag |
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4.2.4.3 Protease Cleavage -- 4.2.4.4 Co-Plasmid -- 4.3 Choosing an Appropriate Host -- Exercise -- Further Reading -- 5: Gene Cloning -- 5.1 Gene Amplification from Genomic DNA -- 5.1.1 Ordering a Bacterial Strain or Its Genomic DNA -- 5.1.2 Bacterial Cultivation and Genomic DNA Extraction -- 5.1.3 Gene Amplification -- 5.1.3.1 Primer Design and Ordering Primers -- 5.1.3.2 Polymerase Chain Reaction (PCR) and DNA Polymerases -- 5.2 Gene Synthesis -- 5.2.1 Gene Design -- 5.2.2 Ordering a Synthetic Gene and Codon Optimization -- 5.3 Molecular Cloning -- 5.3.1 Restriction and Ligation |
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5.3.2 NEBuilder® HiFi DNA Assembly -- 5.3.3 PTO-QuickStep -- 5.4 Clone Verification -- Exercise -- Further Reading -- 6: Protein Expression -- 6.1 Protein Expression Medium -- 6.2 Protein Expression Conditions -- 6.3 Inclusion Body -- Exercise -- Further Reading -- 7: Assay -- 7.1 Assay Detection Mode and Formats -- 7.1.1 Assay Detection Mode -- 7.1.1.1 Absorbance -- 7.1.1.2 Fluorescence -- 7.1.1.3 Others -- 7.1.2 Assay Formats -- 7.1.2.1 Microplate -- 7.1.2.2 Agar Plate, Membrane and Digital Imaging -- 7.1.2.3 Paper-Based Assay -- 7.1.2.4 Fluorescence-Activated Cell Sorting (FACS) |
| Summary |
This textbook introduces readers in an accessible and engaging way to the nuts and bolts of protein expression and engineering. Various case studies illustrate each step from the early sequence searches in online databases over plasmid design and molecular cloning techniques to protein purification and characterization. Furthermore, readers are provided with practical tips to successfully pursue a career as a protein engineer. With protein engineering being a fundamental technique in almost all molecular biology labs, the book targets advanced undergraduates and graduate students working in molecular biology, biotechnology and related scientific fields |
| Bibliography |
Includes bibliographical references |
| Notes |
Online resource; title from digital title page (viewed on December 29, 2020) |
| Subject |
Protein engineering.
|
|
Biochemical engineering.
|
|
Protein Engineering
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|
Biochemical engineering
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Protein engineering
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| Form |
Electronic book
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| Author |
Tee, Kang Lan, author
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| ISBN |
9783030568986 |
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3030568989 |
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