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This volume provides state-of-the-art and novel methods on antibiotic isolation and purification, identification of antimicrobial killing mechanisms, and methods for the analysis and detection of microbial adaptation strategies. Antibiotics: Methods and Protocols guides readers through chapters on production and design, mode of action, and response and susceptibility. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Antibiotics: Methods and Protocols aims to inspire scientific work in the exciting field of antibiotic research.

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Part I: Production and Design

1. Antibiotics - Precious Goods in Changing Times

Peter Sass

2. Mining Bacterial Genomes for Secondary Metabolite Gene Clusters

Martina Adamek, Marius Spohn, Evi Stegmann, and Nadine Ziemert

3. Production of Antimicrobial Compounds by Fermentation

Henrik Harms, Gabriele M. König and Till F. Schäberle

4. Structure Elucidation of Antibiotics by NMR Spectroscopy

Georgios Daletos, Elena Ancheeva, Raha Orfali, Victor Wray, and Peter Proksch

5. Computer-Aided Drug Design Methods

Wenbo Yu and Alexander D. MacKerell, Jr.

6. Cytotoxicity Assays as Predictors of the Safety and Efficacy of Antimicrobial Agents

Alexander Zipperer and Dorothee Kretschmer

Part II: Mode of Action

7. Application of a Bacillus subtilis Whole-cell Biosensor ( P liaI -lux ) for the Identification of Cell Wall Active Antibacterial Compounds

Carolin Martina Kobras, Thorsten Mascher, and Susanne Gebhard

8. Determination of Bacterial Membrane Impairment by Antimicrobial Agents

Miriam Wilmes and Hans-Georg Sahl

9. Mass-sensitive Biosensor Systems to Determine the Membrane Interaction of Analytes

Sebastian G. Hoß and Gerd Bendas

10. Measurement of Cell Membrane Fluidity by Laurdan GP:Fluorescence Spectroscopy and Microscopy

Kathi Scheinpflug, Oxana Krylova and Henrik Strahl

11. In vitro Assays to Identify Antibiotics Targeting DNA Metabolism
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12. Fluorescence-based real-time Activity Assays to Identify RNase P inhibitors

Yu Chen, Xin Liu, Nancy Wu, and Carol A. Fierke

13. Reporter Gene-based Screening for TPP Riboswitch Activators

Christina E. Lünse and Günter Mayer

14. Cell-based Fluorescent Screen to Identify Inhibitors of Bacterial Translation Initiation
Federica Briani

15. Bacterial Histidine Kinases: Overexpression, Purification, and Inhibitor Screen

Mike Gajdiss, Michael Türck, and Gabriele Bierbaum

Part III: Response and Susceptibility

16. Expression Profiling of Antibiotic Resistant Bacteria Obtained by Laboratory Evolution

Shingo Suzuki, Takaaki Horinouchi, and Chikara Furusawa

17. Sample Preparation for Mass-Spectrometry Based Absolute Protein Quantification in Antibiotic Stress Research
Florian Bonn, Sandra Maass, and Dörte Becher

18. Label-Free Quantitation of Ribosomal Proteins from Bacillus Subtillis for Antibiotic Research

Sina Schäkermann, Pascal Prochnow, and Julia E. Bandow

19. Functional Metagenomics to Study Antibiotic Resistance

Manish Boolchandani, Sanket Patel, and Gautam Dantas

20. Epidemiological Surveillance and Typing Methods to Track Antibiotic Resistant Strains using High Throughput Sequencing

Miguel Paulo Machado, Bruno Ribeiro-Gon‡alves, Mickael Silva, Mário Ramirez, JoÆo André Carri‡o

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