The burgeoning field of genetic code expansion provides new tools for specifically labeling proteins for a variety of applications. Clickable non-canonical amino acids (ncAA) have been refined for almost-quantitative and highly selective reactions with complementary probes. In my thesis work I have adapted genetic code expansion for two biological questions. First, I used ncAA incorporation and click reaction to test the effect of fluorescent protein (FP) tagging on the nanoscale organization of target proteins. Second, I used these tools to generate a genetically-encoded scheme for specifi...

This detailed volume explores non-canonical amino acids (ncAAs) through their site-specific incorporation by genetic code expansion (GCE). The collection provides a broad resource of methods for implementing GCE in E. coli, mammalian cells, and animals, highlighting specific applications ranging from fluorescence labeling to photocontrol and the study of protein post-translational modification. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Genetically Incorporated Non-Canonical Amino Acids: Methods and Protocols serves as an ideal source of methodologies that can be adapted and extended, migrated to different model systems, and combined in new ways to help explore a wide range of biological questions and to augment industrial and pharmaceutical protein engineering.

By combining the tools of organic chemistry with those of physical biochemistry and cell biology, Non-Natural Amino Acids aims to provide fundamental insights into how proteins work within the context of complex biological systems of biomedical interest. The critically acclaimed laboratory standard for 40 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. With more than 400 volumes published, each Methods in Enzymology volume presents material that is relevant in today's labs -- truly an essential publication for researchers in all fields of life sciences. - Demonstrates how the tools and principles of chemistry combined with the molecules and processes of living cells can be combined to create molecules with new properties and functions found neither in nature nor in the test tube - Presents new insights into the molecular mechanisms of complex biological and chemical systems that can be gained by studying the structure and function of non-natural molecules - Provides a "one-stop shop" for tried and tested essential techniques, eliminating the need to wade through untested or unreliable methods

Stimulated Raman Scattering Microscopy: Techniques and Applications describes innovations in instrumentation, data science, chemical probe development, and various applications enabled by a state-of-the-art stimulated Raman scattering (SRS) microscope. Beginning by introducing the history of SRS, this book is composed of seven parts in depth including instrumentation strategies that have pushed the physical limits of SRS microscopy, vibrational probes (which increased the SRS imaging functionality), data science methods, and recent efforts in miniaturization. This rapidly growing field needs a comprehensive resource that brings together the current knowledge on the topic, and this book does just that. Researchers who need to know the requirements for all aspects of the instrumentation as well as the requirements of different imaging applications (such as different types of biological tissue) will benefit enormously from the examples of successful demonstrations of SRS imaging in the book. Led by Editor-in-Chief Ji-Xin Cheng, a pioneer in coherent Raman scattering microscopy, the editorial team has brought together various experts on each aspect of SRS imaging from around the world to provide an authoritative guide to this increasingly important imaging technique. This book is a comprehensive reference for researchers, faculty, postdoctoral researchers, and engineers. - Includes every aspect from theoretic reviews of SRS spectroscopy to innovations in instrumentation and current applications of SRS microscopy - Provides copious visual elements that illustrate key information, such as SRS images of various biological samples and instrument diagrams and schematics - Edited by leading experts of SRS microscopy, with each chapter written by experts in their given topics

This volume aims to provide a timely view of the state-of-the-art in systems biology. The editors take the opportunity to define systems biology as they and the contributing authors see it, and this will lay the groundwork for future studies. The volume is well-suited to both students and researchers interested in the methods of systems biology. Although the focus is on plant systems biology, the proposed material could be suitably applied to any organism.

Memory Mechanisms is an edited review volume that summarizes state-of-the-art knowledge on memory mechanisms at the molecular, cellular and circuit level. Each review is written by leading experts in the field, presenting not only current knowledge, but also discussing the concepts, providing critical reflections and suggesting an outlook for future studies. The memory mechanisms are also discussed in the context of diseases. Studies of memory deficits in disease models are introduced as well as approaches to restore memory deficits. Finally, the impact of contemporary memory research for psychiatry is illustrated.

The ability to introduce non-canonical amino acids in vivo has greatly expanded the repertoire of accessible proteins for basic research and biotechnological application. Here, the different methods and strategies to incorporate new or modified amino acids are explained in detail, including a lot of practical advice for first-time users of this powerful technique. Novel applications in protein biochemistry, genomics, biotechnology and biomedicine made possible by the expansion of the genetic code are discussed and numerous examples are given. Essential reading for all molecular life scientists who want to stay ahead in their research.