Chapter
Publicly Available
13 Determining the Efficiency of the Enzyme Acetylcholine Esterase Using Steady-State Kinetic Experiment
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Timea Gerczei
Chapters in this book
- Frontmatter I
- Preface V
- Contents IX
- 1 Introducing the Bacterial Antibiotic Sensor Mini Project 1
- 2 Identifying Conserved Elements in the Toxin Sensor and Designing Mutants to Test Whether They are Important for Function 9
- 3 Designing Primers for Site-Directed Mutagenesis 27
- 4 Performing Site-Directed Mutagenesis 38
- 5 Purifying Mutant Toxin Sensor DNA from Bacterial Cells and Evaluating its Quality Using Agarose Gel Electrophoresis and UV Spectroscopy 51
- 6 Preparing DNA Template for Mutant RNA Sensor Synthesis Using a Restriction Endonuclease 64
- 7 Synthesizing the ykkCD Mutant Toxin Sensor RNA in vitro 81
- 8 Purifying the ykkCD Mutant Toxin Sensor RNA and Evaluating its Purity Using Denaturing PAGE and UV spectrometry 89
- 9 Evaluating the Ability of the ykkCD Toxin Sensor to Recognize the Antibiotic Tetracycline Using Fluorescent Quenching 97
- 10 Evaluating Antibiotic Binding to Blood Serum Albumin Using Fluorescence Spectroscopy 113
- 11 Understanding the Importance of Buffers in Biological Systems 124
- 12 Molecular Visualization of an Enzyme, Acetylcholinesterase 132
- 13 Determining the Efficiency of the Enzyme Acetylcholine Esterase Using Steady-State Kinetic Experiment 146
- 14 Separation of the Phosphatidylcholines Using Reverse Phase HPLC 161
- List of Figures 171
Chapters in this book
- Frontmatter I
- Preface V
- Contents IX
- 1 Introducing the Bacterial Antibiotic Sensor Mini Project 1
- 2 Identifying Conserved Elements in the Toxin Sensor and Designing Mutants to Test Whether They are Important for Function 9
- 3 Designing Primers for Site-Directed Mutagenesis 27
- 4 Performing Site-Directed Mutagenesis 38
- 5 Purifying Mutant Toxin Sensor DNA from Bacterial Cells and Evaluating its Quality Using Agarose Gel Electrophoresis and UV Spectroscopy 51
- 6 Preparing DNA Template for Mutant RNA Sensor Synthesis Using a Restriction Endonuclease 64
- 7 Synthesizing the ykkCD Mutant Toxin Sensor RNA in vitro 81
- 8 Purifying the ykkCD Mutant Toxin Sensor RNA and Evaluating its Purity Using Denaturing PAGE and UV spectrometry 89
- 9 Evaluating the Ability of the ykkCD Toxin Sensor to Recognize the Antibiotic Tetracycline Using Fluorescent Quenching 97
- 10 Evaluating Antibiotic Binding to Blood Serum Albumin Using Fluorescence Spectroscopy 113
- 11 Understanding the Importance of Buffers in Biological Systems 124
- 12 Molecular Visualization of an Enzyme, Acetylcholinesterase 132
- 13 Determining the Efficiency of the Enzyme Acetylcholine Esterase Using Steady-State Kinetic Experiment 146
- 14 Separation of the Phosphatidylcholines Using Reverse Phase HPLC 161
- List of Figures 171