Cells possess a wealth of posttranscriptional control mechanisms that impact on every conceivable aspect of the life of an mRNA. These processes are intimately intertwined in an almost baroque manner, where promoter context influences the recruitment of splicing factors, where the majority of pre-mRNAs undergo alternative splicing, and where proteins deposited during nuclear processing impact distal cytoplasmic processing, translation, and decay. If there is a unifying theme to mRNA Processing and Metabolism: Methods and Protocols, it is that mRNA processing and metabolism are integrated...

Leading scientists in gene expression methodology and bioinformatics data analysis describe readily reproducible methods for measuring RNA levels in cells and tissues. The techniques presented include new methods for applying the Affymetrix GeneChip(r), SAR-SAGE, StaRT-PCR, SSH, the Invader Assay(r), and ADGEM. The authors also provide critical bioinformatics insight and resources for data analysis and management. By distilling the basic underlying principles of many methods to a few straightforward concepts, investigators can easily choose the method most appropriate to their application

Gazing into crystal balls is beyond the expertise of most scientists. Yet, as we look further into the 21st century, one does not have to be Nostradamus to predict that the current genomics and proteomics "revolution" will have an immense impact on medical bacteriology. This impact is already being re- ized in many academic departments, and although encroachment on routine diagnostic bacteriology, particularly in the hospital setting, is likely to occur at a slower pace, it remains nonetheless inevitable. Therefore, it is important that no one working in bacteriology should find themselves...

A diverse collection of state-of-the-art methods for the microscopic imaging of cells and molecules. The authors cover a wide spectrum of complimentary techniques, including such methods as fluorescence microscopy, electron microscopy, atomic force microscopy, and laser scanning cytometry. Additional readily reproducible protocols on confocal scanning laser microscopy, quantitative computer-assisted image analysis, laser-capture microdissection, microarray image scanning, near-field scanning optical microscopy, and reflection contrast microscopy round out this eclectic collection of...

A comprehensive collection of readily reproducible techniques for the manipulation of recombinant plasmids using the bacterial host E. coli. The authors describe proven methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and analyzing recombinant clones. They also include protocols for the construction and screening of libraries, as well as specific techniques for specialized cloning vehicles, such as cosmids, bacterial artificial chromosomes, l vectors, and phagemids. Common downstream applications such as mutagenesis of plasmids, recombinant protein expression,...

Functional genomics is a young discipline whose origin can be traced back to the late 1980s and early 1990s, when molecular tools became available to determine the cellular functions of genes. Today, functional genomics is p- ceived as the analysis, often large-scale, that bridges the structure and organi- tion of genomes and the assessment of gene function. The completion in 2000 of the genome sequence of Arabidopsis thaliana has created a number of new and exciting challenges in plant functional genomics. The immediate task for the plant biology community is to establish the functions of...

The fundamental question of how cells grow and divide has perplexed biologists since the development of the cell theory in the mid-19th century, when it was recognized by Virchow and others that all cells come from cells. In recent years, considerable effort has been applied to the identification of the basic molecules and mechanisms that regulate the cell cycle in a number of different organisms. Such studies have led to the elucidation of the central paradigms that underpin eukaryotic cell cycle control, for which Lee Hartwell, Tim Hunt, and Paul Nurse were jointly awarded the Nobel Prize...

Since the initial discovery of the G protein-coupled receptor system that regulates cyclicAMP production, the G protein field has rapidly expanded. Cell surface receptors that couple to heterotrimeric G proteins, the G prote- coupled receptors (GPCRs), number in the hundreds and bind to a wide div- sity of ligands including, biogenic amines (e. g., adrenaline), lipid derivatives (e. g., lysophosphatidic acid), peptides (e. g., opioid peptides), proteins (e. g., thyroid-stimulating hormone), and odorants to name a few. The GPCR system is found throughout biology in such simple organisms as...

Knowledge of the three-dimensional structure of a protein is absolutely required for the complete understanding of its function. The spatial orientation of amino acids in the active site of an enzyme demonstrates how substrate specificity is defined, and assists the medicinal chemist in the design of s- cific, tight-binding inhibitors. The shape and contour of a protein surface hints at its interaction with other proteins and with its environment. Structural ana- sis of multiprotein complexes helps to define the role and interaction of each individual component, and can predict the...

As the mysteries stored in our DNA have been more completely revealed, scientists have begun to face the extraordinary challenge of unraveling the int- cate network of protein protein interactions established by that DNA fra- work. It is increasingly clear that proteins continuously interact with one another in a highly regulated fashion to determine cell fate, such as proliferation, diff- entiation, or death. These protein protein interactions enable and exert str- gent control over DNA replication, RNA transcription, protein translation, macromolecular assembly and degradation, and signal...