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Transgenic Plants
The aim of Transgenic Plants: Methods and Protocols is to provide a source of information to guide the reader through a wide range of frequently used, broadly applicable, and easily reproducible techniques involved in the gene- tion of transgenic plants. Its step-by-step approach covers a series of methods for genetically transforming plant cells and tissues, and for recovering whole transgenic plants from them. The volume then moves on to the use of sele- able and reporter markers, positive selection, marker elimination after rec- ery of transgenic plants, and the analysis of transgene integration, expression, and localization in the plant genome. Although contributors usually refer to mode...
The Peña-Lara Story
After discovering additional information pertaining to his paternal side of the family, author Christopher G. Pea revised his original book to provide the reader with a richly detailed account of each member of the Pea-Lara family, along with their respective spouses. Both highly informative and engaging, The Pea-Lara Story: Revisited retraces the familys roots that began in New Spain (Mexico), including the military exploits of the familys patriarch, Lt. Col. Jos Emeterio Pozas, who served under Spain and Mexico. In addition, the story includes an account of the familys life in Monterrey, Nuevo Len, Mexico, the Pea-Laras forced evacuation of the city during the height of the 1910-1920 Mexic...
Closteroviridae
Plant viruses grouped within this family have remarkable properties, prominent among which is their genomic size: Citrus tristeza virus (CTV) has the largest (19.3 kb) genome reported for a plant monopartite single-stranded RNA (+) virus. Virions are filamentous and typically flexuous particles, approximately 12 nm in diameter and 650 to 2000 nm in length, with a unique bipolar (“rattlesnake”) morphology: the major coat protein (CP) encapsidates most of the genomic RNA, with a minor CP (CPm) coating a small 5’-terminal fragment (virion tail) and other viral-encoded proteins being also incorporated to this tail. The genome is monopartite (genus Closterovirus, type member Beet yellows vi...
Oligonucleotide Synthesis
A collection of powerful new techniques for oligonucleotide synthesis and for the use of modified oligonucleotides in biotechnology. Among the protocol highlights are a novel two-step process that yields a high purity, less costly, DNA, the synthesis of phosphorothioates using new sulfur transfer agents, the synthesis of LNA, peptide conjugation methods to improve cellular delivery and cell-specific targeting, and triple helix formation. The applications include using molecular beacons to monitor the PCR amplification process, nuclease footprinting to study the sequence-selective binding of small molecules of DNA, nucleic acid libraries, and the use of small interference RNA (siRNA) as an inhibitor of gene expression.
RNA Silencing
A collection of readily reproducible methods for the design, preparation, and use of RNAs for silencing gene expression in cells and organisms. The techniques range widely and include methods addressing the biochemical aspects of the silencing machinery, RNA silencing in non-mammalian organisms, and the in vivo delivery of siRNAs and silencing vectors. There are also techniques for designing, preparing, and using RNAs to silence gene expression, for fine-tuning regulation by targeting specific isoforms of a given gene, and for the study and use of microRNAs. The protocols follow the successful Methods in Molecular BiologyTM series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Transgenic Horticultural Crops
As the world debates the risks and benefits of plant biotechnology, the proportion of the global area of transgenic field crops has increased every year, and the safety and value continues to be demonstrated. Yet, despite the success of transgenic field crops, the commercialization of transgenic horticultural crops (vegetables, fruits, nuts, and or
Cell Cycle Control and Dysregulation Protocols
Cell Cycle Control and Dysregulation Protocols focuses on emerging methodologies for studying the cell cycle, kinases, and kinase inhibitors. It addresses the issue of gene expression in vivo and in vitro, the analysis of cyclin-dependent kinase inhibitors, protein degradation mediated by the proteosome, the analysis of the transformed cell phenotype, and innovative techniques to detect apoptosis. Because there are already many manuals and protocols available, along with commercial kits and reagents, a variety of the more common techniques have not been included in our book. The protocols described, based on rather sophisticated techniques for in vivo and in vitro studies, consist of molecular biology, biochemistry, and various types of immunoassays. Indeed, the authors have successfully accomplished an arduous task by presenting several topics in the simplest possible manner. We are confident that Cell Cycle Control and Dysregulation Protocols will facilitate and optimize the work of practical scientists involved in researching the cell cycle. We greatly acknowledge the extraordinary contribution of the authors in writing this book.
Molecular Toxicology Protocols
A collection of cutting-edge techniques for analyzing genotoxic exposure and detecting the resulting biological effects-including endogenous metabolites-up to and including the development of cancer. The authors emphasize analytical methods that can be specifically applied to human populations and patients. Among the applications detailed are the analysis of interactions between such cellular macromolecules as DNA and proteins and chemical and physical agents, the assessment of medically relevant toxicity, and the characterization of genetic alterations induced in transgenic animals by in vivo systems. There are also methods for the analysis of genotoxic exposure during gene expression, of cytotoxicity caused by the induction of apoptosis, of genetic alterations in reporter genes and oncogenes, early (premalignant) detection of altered oncogenes, and of individual variation in biotransformation and DNA repair capacity.
Cell Migration
A collection of classic, novel, and state-of-the-art methods for the study of cell migration in cultured cells, different model organisms, and specialized cells in normal development and disease. Highlights include basic assays that apply to all cell migration studies in vitro, assays in various model organisms, and assays for cancer cells, endothelial cells, and neurons both in vitro and in animal models. The protocols follow the successful Methods in Molecular BiologyTM series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
