Synopses & Reviews
Parasitic disease remains an important health issue in most regions of the world and constitutes a major cause of morbidity and mortality in developing countries. In Parasite Genomics Protocols, pioneering bench researchers describe in detail the cutting-edge techniques they have developed for analyzing the genomes and gene products in a diverse range of protozoan and metazoan parasites. These readily reproducible techniques can be used in genomic, functional genomic, and postgenomic studies and include transfection methods and vectors for several protozoan parasites, global analysis using microarrays, gene ablation using RNA interference, gene knockout, mutagenesis, and chromosome manipulation. Some of the protocols require DNA sequence data, whereas others were developed independently of whole-genome sequence data. Additional chapters survey the available genome sequences, helping novice researchers overcome the difficulties of accessing the databases and applying bioinformatics analysis to biological function. The protocols presented follow the successful Methods in Molecular Biology™ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and the avoidance of known pitfalls. State-of-the-art and highly practical, Parasite Genomics Protocols offers bench investigators in parasitology a wide-ranging collection of diverse methods for elucidating the content of the genomes of parasitic organisms, utilizing the genome sequence for genetic analysis, and ascribing function to the available genome sequences.
Review
"...represents an excellent resource for both students and researchers in the field." - Society for General Microbiolgy
Synopsis
Parasitic diseases remain a major health problem throughout the world, for both humans and animals. For many of us, our technologically advanced lifestyle has decreased the prevalence and transmission of parasitic diseases, but for the majority of the world's population, they are ever present in homes, domestic animals, food, or the environment. The study of parasites and parasitic disease has a long and distinguished history. In some cases, it has been driven by the great importance of the presence of the parasite to the community, for example, those that affect our livestock. In other cases, it is clear that applied research has suffered for lack of funding because the parasite affects people with few resources, such as the rural poor in resource-poor countries. These instances include the so-called "neglected diseases," as defined by the World Health Organization (WHO). Parasites have complicated life cycles, and a thorough understanding of the unique characteristics of a particular parasite species is vital in attempts to avoid, prevent, or cure infection or to alleviate symptoms. Of course, the biological characteristics that each parasite has developed to aid survival and transmission, to avoid destruction by the immune system, and to adapt to a changing environment are of lasting fascination to basic biologists as well. The elegance of these biological systems has ensured that the study of protozoan and metazoan parasites also remains an active field of research in countries where the diseases are not a threat to the population.
Synopsis
Pioneering bench researchers describe in detail the cutting-edge techniques they have developed for analyzing the genomes and gene products in a diverse range of protozoan and metazoan parasites. These readily reproducible techniques can be used in genomic, functional genomic, and post-genomic studies and include transfection methods and vectors for several protozoan parasites, global analysis using microarrays, gene ablation using RNA interference, gene knockout, mutagenesis, and chromosome manipulation. Some of the protocols require DNA sequence data, whereas others were developed independently of whole-genome sequence data. The protocols presented follow the successful Methods in Molecular Biology™ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and the avoidance of known pitfalls.
Table of Contents
Sequencing Strategies for Parasite Genomes Daniella Bartholomeu and Najib M. El-Sayed Annotation of Parasite Genomes Matthew Berriman and Midori Harris Parasite Genome Databases and Web-Based Resources Christiane Hertz-Fowler and Neil Hall Expressed Sequence Tags: Medium-Throughput Protocols Claire Whitton, Jennifer Daub, Marian Thompson, and Mark Blaxter Expressed Sequence Tags: Analysis and Annotation John Parkinson and Mark Blaxter Positive Selection Scanning of Parasite DNA Sequences Winston A. Hide and Raphael D. Isokpehi RACE and RAGE Cloning in Parasitic Microbial Eukaryotes Bryony A. P. Williams and Robert P. Hirt Amplified (Restriction) Fragment Length Polymorphism (AFLP) Analysis Daniel K. Masiga and C. Michael R. Turner Minisatellites and MVR-PCR for the Individual Identification of Parasite Isolates Annette MacLeod Analysis of Differentially Expressed Parasite Genes and Proteins Using Transcriptomics and Proteomics Daniel C. Gare Gene Expression Studies Using Self-Fabricated Parasite cDNA Microarrays Karl F. Hoffmann and Jennifer M. Fitzpatrick DNA Content Analysis on Microarrays Upinder Singh, Preetam H. Shah, and Ryan C. MacFarlane Typing Single-Nucleotide Polymorphisms in Toxoplasma gondii by Allele-Specific Primer Extension and Microarray Detection Chunlei Su, Christian Hott, Bernard H. Brownstein, and L. David Sibley Transfection of the Human Malaria Parasite Plasmodium falciparum Brendan S. Crabb, Melanie Rug, Tim-Wolf Gilberger, Jennifer K. Thompson, Tony Triglia, Alexander G. Maier, and Alan F. Cowman A PCR-Based Method for Gene Deletion and Protein Tagging in Trypanosoma brucei George K. Arhin, Shuiyuan Shen, Elisabetta Ullu, and Christian Tschudi Analysis of Gene Function in Trypanosoma brucei Using RNA Interference Appolinaire Djikeng, Shuiyuan Shen, Christian Tschudi, and Elisabetta Ullu In Vitro Shuttle Mutagenesis Using Engineered Mariner Transposons Kelly A. Robinson, Sophie Goyard, and Stephen M. Beverley Random Mutagenesis Strategies for Construction of Large and Diverse Clone Libraries of Mutated DNA Fragments Sudsanguan Chusacultanachai and Yongyuth Yuthavong Separation, Digestion, and Cloning of Intact Parasite Chromosomes Embedded in Agarose Vanessa Leech, Michael A. Quail, and Sara E. Melville Chromosome Fragmentation in Leishmania Pascal Dubessay, Christine Blaineau, Patrick Bastien, and Michel Pagès FISH Mapping for Helminth Genomes Hirohisa Hirai and Yuriko Hirai Fiber-FISH: Fluorescence In Situ Hybridization on Stretched DNA Klaus Ersfeld Yeast Two-Hybrid Assay for Studying Protein-Protein Interactions Ahmed Osman From Genomes to Vaccines for Leishmaniasis Carmel B. Stober Index