Protocols in Molecular Parasitology

ISBN-13: 9780896032392 / Angielski / Miękka / 1993 / 488 str.

John E. Hyde

Opis

Protocols in Molecular Parasitology offers a wide range of experi- mental protocols, each written by experts, to research workers interested in exploring the molecular aspects of parasitology. This is a field that has expanded enormously in scope and potential over the last decade, although it is as yet significantly underrepresented in the literature out- side of academic journals. The need for manuals such as this one stems in part from the increasing difficulty of reproducing experimental tech- niques from reports in standard journal papers, where pressures on space usually result in highly abbreviated descriptions. The "Notes" sections of the following chapters are particularly aimed at counteracting this problem. Since the emphasis of the book is on aspects of molecular analysis, it was possible to devote only a limited amount of space to describing protocols for culture and isolation of the primary parasitic material. Therefore, of the numerous parasitic systems that are currently being investigated in laboratories around the world, only techniques for c- turing the "big five" are described here, since they collectively account for the great bulk of research in this field. It is also possible that some minor front-end modifications to certain protocols may be necessary, depending on exactly which parasite is being studied.

Recenzje

...well-written and thoughtfully assembled...will be welcomed by recent arrivals...and regularly utilized by instructors of future investigators. - Analytical Biochemistry

Spis treści

Culturing and Biological Cloning of T. brucei. Culturing and Biological Cloning of T. cruzi. In Vitro Cultivation and Biological Cloning of Leishmania. Simple In Vitro Cultivation of the Malaria Parasite P. falciparum (Erythrocytic Stages) Suitable for Large-Scale Preparations. Synchronization and Cloning of Malaria Parasites. The Culture and Prparation of Gametocytes of Plasmodium falciparum for Immunochemical, Molecular, and Mosquito Infectivity Studies. The Culture of S. mansoni and Production of Life Cycle Stages. Preparation of DNA and RNA from T. brucei. Isolation of RNA and DNA from T. cruzi. Isolation of DNA and RNA from Leishmania. The Extraction and Purification of DNA and RNA from In Vitro Cultures of the Malaria Parasite P. falciparum. Extraction and Purificaiton of Nucleic Acids from Schistosomes. Application of DNA and Whole Organisms to Filter Supports for DNA Probe Analysis: Dot, Slot, and Touch Blotting. The Development and Use of Repetitive Sequences as DNA Probes for Parasite Detection and Species Identification. Analysis of the DNA of Parasitic Protozoa by Flow Cytometry. PCR for Low-Level Detection of Malaria Parasites in Blood. PCR Methods for Identification of Point Mutations and Gene Rearrangements. Antigenic Typing of Field Isolates of Plasmodium by DNA Techniques. Ribosomal RNA Probes for Detection and Identification of Species. Rapid Sequencing of Parasite rRNA. Antibody Screening of Expression Libraries. Antibody Select Procedure for Characterization of Expression Clones. Isolation of Parasite Genes Using Synthetic Oligonucleotides. Chromosome Mapping Methods for Parasitic Protozoa. Transfection of Leishmania and Trypanosoma brucei by Electroporation. Two-Dimensional Polyacrylamide Gel Electrophoresis. Isoenzyme Electrophoresis for Parasiste Characterization. The Surface Labeling of Schistosomes. Monoclonal Antibody Affinity Chromatography. Immunoblotting and Enzyme Linked Immunosorbent Assay. Immunofluorescence of Parasites. Immunoelectron Microscopic Localization of Antigens in Malaria Parasites. Preparation of Blotted Membrane for Protein Microsequencing. Identification of T-Cell Epitopes. Index.