Cell Cycle Checkpoint Control Protocols

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Cell Cycle Checkpoint Control Protocols

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Author : Howard B. Lieberman
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-02

Cell Cycle Checkpoint Control Protocols written by Howard B. Lieberman and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-02 with Science categories.

The field of cell cycle regulation is based on the observation that the life cycle of a cell progresses through several distinct phases, G1, M, S, and G2, occurring in a well-defined temporal order. Details of the mechanisms involved are rapidly emerging and appear extraordinarily complex. Furthermore, not only is the order of the phases important, but in normal eukaryotic cells one phase will not begin unless the prior phase is completed successfully. Che- point control mechanisms are essentially surveillance systems that monitor the events in each phase, and assure that the cell does not progress prematurely to the next phase. If conditions are such that the cell is not ready to progress—for example, because of incomplete DNA replication in S or DNA damage that may interfere with chromosome segregation in M—a transient delay in cell cycle progression will occur. Once the inducing event is properly handled— for example, DNA replication is no longer blocked or damaged DNA is repaired—cell cycle progression continues. Checkpoint controls have recently been the focus of intense study by investigators interested in mechanisms that regulate the cell cycle. Furthermore, the relationship between checkpoint c- trol and carcinogenesis has additionally enhanced interest in these cell cycle regulatory pathways. It is clear that cancer cells often lack these checkpoints and exhibit genomic instability as a result. Moreover, several tumor suppressor genes participate in checkpoint control, and alterations in these genes are as- ciated with genomic instability as well as the development of cancer.

Directed Enzyme Evolution

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Author : Frances H. Arnold
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-02

Directed Enzyme Evolution written by Frances H. Arnold and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-02 with Science categories.

Directed evolution comprises two distinct steps that are typically applied in an iterative fashion: (1) generating molecular diversity and (2) finding among the ensemble of mutant sequences those proteins that perform the desired fu- tion according to the specified criteria. In many ways, the second step is the most challenging. No matter how cleverly designed or diverse the starting library, without an effective screening strategy the ability to isolate useful clones is severely diminished. The best screens are (1) high throughput, to increase the likelihood that useful clones will be found; (2) sufficiently sen- tive (i. e. , good signal to noise) to allow the isolation of lower activity clones early in evolution; (3) sufficiently reproducible to allow one to find small improvements; (4) robust, which means that the signal afforded by active clones is not dependent on difficult-to-control environmental variables; and, most importantly, (5) sensitive to the desired function. Regarding this last point, almost anyone who has attempted a directed evolution experiment has learned firsthand the truth of the dictum “you get what you screen for. ” The protocols in Directed Enzyme Evolution describe a series of detailed p- cedures of proven utility for directed evolution purposes. The volume begins with several selection strategies for enzyme evolution and continues with assay methods that can be used to screen enzyme libraries. Genetic selections offer the advantage that functional proteins can be isolated from very large libraries s- ply by growing a population of cells under selective conditions.

Biopolymer Methods In Tissue Engineering

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Author : Anthony P. Hollander
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-03

Biopolymer Methods In Tissue Engineering written by Anthony P. Hollander and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-03 with Technology & Engineering categories.

There is an urgent need to develop new approaches to treat conditions as- ciated with the aging global population. The surgeon’s approach to many of these problems could be described as having evolved through three stages: Removal: Traditionally, diseased or badly damaged tissues and structures might simply be removed. This was appropriate for limbs and non-essential organs, but could not be applied to structures that were critical to sustain life. An additional problem was the creation of disability or physical deformity that in turn could lead to further complications. Replacement: In an effort to treat wider clinical problems, or to overcome the limitations of amputation, surgeons turned to the use of implanted materials and medical devices that could replace the functions of biological structures. This field developed rapidly in the 1960s and 1970s, with heart valve and total joint replacement becoming common. The term “biomaterial” was used increasingly to describe the materials used in these operations, and the study of biomaterials became one of the first truly interdisciplinary research fields. Today, biomaterials are employed in many millions of clinical procedures each year and they have become the mainstay of a very successful industry.

Gene Delivery To Mammalian Cells

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Author : William C. Heiser
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-03

Gene Delivery To Mammalian Cells written by William C. Heiser and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-03 with Science categories.

Experienced researchers describe in step-by-step detail methods that have proven most useful in delivering genes to mammalian cells. Volume 1 focuses on gene delivery by a variety of chemical and physical methods, including ultrasound, biolistics, peptides, PNA clamps, liposomes, microinjection, electroporation, particle bombardment, dendrimers, and hydrodynamics. Volume 2 details procedures for delivering genes to cells in vitro and in vivo, including the use of lentiviral vectors.

Cell Cycle Checkpoints

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Author : Willis X. Li
language : en
Publisher: Humana Press
Release Date : 2011-08-31

Cell Cycle Checkpoints written by Willis X. Li and has been published by Humana Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2011-08-31 with Science categories.

Cell cycle checkpoints control the fidelity and orderly progression of eukaryotic cell division. By controlling the orderly progression of critical cell cycle events such as DNA replication and chromosome segregation and ensuring proper repair of damaged DNA, cell cycle checkpoints function to ensure genome integrity. Mechanisms of checkpoint controls are not only the research focus of investigators interested in mechanisms that regulate the cell cycle, but are also the interests of researchers studying cancer development as it is increasingly clear that loss of cell cycle checkpoints, which leads to genomic instability as a result, is a hallmark of tumorigenesis. Cell Cycle Checkpoints: Methods and Protocols provides detailed descriptions of methodologies currently employed by researchers in the field, including those commonly used in the mammalian, yeast, C. elegans, Drosophila, and Xenopus model systems. Each chapter describes a specific technique or protocol, such as a method to induce cell cycle checkpoints in a particular model system, to synchronize a population of cells to allow observations of cell cycle progression, to identify genes involved in checkpoint regulation, and to study particular protein components of cell cycle checkpoint pathways. Written in the highly successful Methods in Molecular BiologyTM series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Cell Cycle Checkpoints: Methods and Protocols seeks to serve both professionals and novices with its well-honed methodologies in an effort to further our knowledge of this essential field.

Human Retrovirus Protocols

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Author : Tuofu Zhu
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-04

Human Retrovirus Protocols written by Tuofu Zhu and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-04 with Science categories.

A cutting-edge collection of basic and state-of-the-art methods optimized for investigating the molecular biology of this class of retrovirus. These readily reproducible techniques range from methods for the isolation and detection of human retroviruses to cutting-edge methods for exploring the interplay between the viruses and the host. Here, the researcher will find up-to-date techniques for the isolation and propagation of HIV, HTLV, and foamy virus from a variety of sources. There are also assays for determining the cell tropism of HIV-1, the coreceptor usage of HIV-1, and human gene expression with HIV-1 infection by microarrays, as well as for phenotyping HIV-1 infected monocytes and examining their fitness. Highlights include the detection and quantification of HIV-1 in resting CD4+, a new cloning system for making recombinent virus, cDNA microarrays, and the determination of genetic polymorphisms in two recently identified HIV-1 co-factors that are critical for HIV-1 infection.

Checkpoint Controls And Cancer

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Author : Axel H. Schönthal
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-03

Checkpoint Controls And Cancer written by Axel H. Schönthal and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-03 with Medical categories.

Intracellular checkpoint controls constitute a network of signal transd- tion pathways that protect cells from external stresses and internal errors. Ext- nal stresses can be generated by the continuous assault of DNA-damaging agents, such as environmental mutagens, ultraviolet (UV) light, ionizing radiation, or the reactive oxygen species that can arise during normal cellular metabolism. In response to any of these assaults on the integrity of the genome, the activation of the network of checkpoint control pathways can lead to diverse cellular responses, such as cell cycle arrest, DNA repair, or elimination of the cell by cell death (apoptosis) if the damage cannot be repaired. Moreover, internal errors can occur during the highly orchestrated replication of the cellular genome and its distribution into daughter cells. Here, the temporal order of these cell cycle events must be strictly enforced—for example, to ensure that DNA replication is c- plete and occurs only once before cell division, or to monitor mitotic spindle assembly, and to prevent exit from mitosis until chromosome segregation has been completed. Thus, well functioning checkpoint mechanisms are central to the maintenance of genomic integrity and the basic viability of cells and, the- fore, are essential for proper development and survival. The importance of proper functioning of checkpoints becomes plainly obvious under conditions in which this control network malfunctions and fails. Depending on the severity and timing, failure of this machinery can lead to embryonic lethality, genetic diseases, and cancer.

Receptor Binding Techniques

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Author : Anthony P. Davenport
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-02

Receptor Binding Techniques written by Anthony P. Davenport and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-02 with Medical categories.

A comprehensive collection of readily reproducible methods for studying receptors in silico, in vitro, and in vivo. These cutting-edge techniques cover mining from curated databases, identifying novel receptors by high throughput screening, molecular methods to identify mRNA encoding receptors, radioligand binding assays and their analysis, quantitative autoradiography, and imaging receptors by positron emission tomography (PET). Highlights include phenotypic characterization of receptors in knockout mice, imaging receptors using green fluorescent protein and fluorescent resonance energy transfer, and quantitative analysis of receptor mRNA by TaqMan PCR. These book equips the researcher with techniques for exploring the unprecedented number of new receptor systems now emerging and the so-called "orphan" receptors whose activating ligand has not been identified.

Peptide Synthesis And Applications

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Author : John Howl
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-02

Peptide Synthesis And Applications written by John Howl and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-02 with Science categories.

Hands-on experts describe in step-by-step detail the key methodologies of contemporary peptide synthesis and illustrate their numerous applications. The techniques presented include protocols for chemical ligation, the synthesis of cyclic and phosphotyrosine-containing peptides, lipoamino acid- and sugar-conjugated peptides, and peptide purification and analyses. Additional chapters detail methodologies and instrumentation for high-throughput peptide synthesis, many different applications of peptides as novel research tools and biological probes, and the design and application of fluorescent substrate-based peptides that can be used to determine the selectivity and activity of peptidases. A practical guide to the identification of proteins using mass spectrometric analyses of peptide mixtures is also included.

Protein Nmr Techniques

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Author : A. Kristina Downing
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-03

Protein Nmr Techniques written by A. Kristina Downing and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-03 with Science categories.

When I was asked to edit the second edition of Protein NMR Techniques, my first thought was that the time was ripe for a new edition. The past several years have seen a surge in the development of novel methods that are truly revolutionizing our ability to characterize biological macromolecules in terms of speed, accuracy, and size limitations. I was particularly excited at the prospect of making these techniques accessible to all NMR labs and for the opportunity to ask the experts to divulge their hints and tips and to write, practically, about the methods. I commissioned 19 chapters with wide scope for Protein NMR Techniques, and the volume has been organized with numerous themes in mind. Chapters 1 and 2 deal with recombinant protein expression using two organisms, E. coli and P. pastoris, that can produce high yields of isotopically labeled protein at a reasonable cost. Staying with the idea of isotopic labeling, Chapter 3 describes methods for perdeuteration and site-specific protonation and is the first of several chapters in the book that is relevant to studies of higher molecular weight systems. A different, but equally powerful, method that uses molecular biology to “edit” the spectrum of a large molecule using segmental labeling is presented in Chapter 4. Having successfully produced a high molecular weight target for study, the next logical step is data acquisition. Hence, the final chapter on this theme, Chapter 5, describes TROSY methods for stru- ural studies.