A major success story of modem molecular biology is the development of technologies to clone and express specific genes. Current applications of recombinant gene products cover a wide spectrum, including gene therapy, production of bioactive pharmaceuticals, synthesis of novel biopolymers, agriculture and animal husbandry, and so on. Inherent in bringing these appli- cations to fruition is the need to design "expression constructs" that will per- mit the ready and specific detection and isolation of the defined recombinant gene products. Recombinant Protein Protocols grows out of the need for...

Ribonucleic acids are central to cellular and molecular processes and perform vital functions in both structural and functional roles. RNA molecules form the bridge between the stable genetic information contained within DNA and enzymes and proteins that carry out much of the metabolism within the cell. Many of the sites of protein synthesis, the ribosomes within the cell, are composed of these ribonucleic acids as are the tRNA molecules that deliver the amino acid building blocks to the ribosomes. Of all the RNA species, the nucleic acid intermediate, messenger RNA, is a desirable source of...

This wide-ranging collection of state-of-the-art techniques for the assay of phospholipid and phospholipid-derived second messengers allows the identification and quantification of signal pathway activation. The assays described cover all the major phospholipases (C, D, A2), as well as sphingomyelinase and associated metabolites. Additional protocols are provided for the assay of phosphoinositide 3-, 4-, and 5-kinase activity and for the separation and identification of phospholipids, diacylglycerol and sphingolipids, as well as their phosphoinositol, choline, and fatty acid metabolites....

Robert Hall and a panel of expert researchers present a comprehensive collection of the most frequently used and broadly applicable techniques for plant cell and tissue culture. Readily reproducible and extensively annotated, the methods cover culture initiation, maintenance, manipulation, application, and long-term storage, with emphasis on techniques for genetic modification and micropropagation. Many of these protocols are currently used in major projects designed to produce improved varieties of important crop plants. Plant Cell Culture Protocols's state-of-the-art techniques are certain...

Inrecent years therehasbeen atremendousincreaseinour understandingofthe functioningofthe cellat the molecular leveL This has been achieved in the main by the invention and developmentof new methodology, particularlyin that area generally referred to as "genetic engineering. " Al- though this revolution has been taking place in the field of nucleic acids research, the protein chemist has at the same timedevelopedfresh methodologytokeeppace with the re- quirements ofpresent-daymolecularbiology. Today's mo- lecularbiologistscannolongerbecontentwithbeingexperts inoneparticulararea alone....

Plant Cell and Tissue Culture continues the high standards of Humana's Methods in Molecular Biology series. Its step-by-step approach (a hallmark of the series) is applied to a wide range of basic laboratory techniques and culture conditions appropriate to plant cells.

Because of the diversity of cell types, species, and culture methods, much of this volume is devoted to the culture of particular cell types and to the regeneration of these cells into whole plants. Special attention is also given to the genetic modification of plants, as well as to the economic significance of plant...

The aim of MHC Protocols is to document protocols that can be used for the analysis of genetic variation within the human major histocompatibility complex (MHC; HLA region). The human MHC encompasses approximately 4 million base pairs on the short arm of chromosome 6 at cytogenetic location 6p21. 3. The region is divided into three subregions. The telomeric class I region contains the genes that encode the HLA class I molecules HLA-A, -B, and -C. The centromeric class II region contains the genes encoding the HLA class II molecules HLA-DR, -DQ, and -DP. In between is the class III region,...

Superantigen Protocols assembles experimental protocols that have proved useful for the study of superantigens. These techniques will allow researchers from various areas of cell biology, microbiology, immunology, biochemistry, and molecular biology to assess the physical characteristics and biological effects of well-known superantigens as well as of putative substances that might have superantigenic activities, and to explore therapies for superantig- induced effects. Microbial exotoxins have been studied for decades as virulence factors because of their pathogenic effects. The term...

Posttranslational Modifications of Proteins: Tools for Functional Proteomics is a compilation of detailed protocols needed to detect and analyze the most important co- and posttranslational modifications of proteins. Though, for reasons of simplicity not explicitly mentioned in the title, both kinds of modifications are covered, whether they occur during, or after, biosynthesis of the protein. My intention was to cover the most significant protein modifications, focusing on the fields of protein function, proteome research, and the characterization of pharmaceutical proteins. The majority of...

Mycotoxins produced by molds are common contaminants of many important crops, including wheat, corn, rice, and peanuts. Some mycotoxins are found in fruits and vegetables. These contaminants have a broad range of toxic effects, including carcinogenicity, neurotoxicity, and reproductive and developmental toxicity. The occurrence of mycotoxins in foods is an unavoidable worldwide problem. About 80 countries have imposed regulatory limits to minimize human and animal exposure to mycotoxins. Regulatory limits, including international standards, have tremendous economic impact and must be...