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This book provides insight into the biology of Wolbachia with a particular focus on methodology. Chapters guide readers through reproductive phenotypes caused by Wolbachia, survival and replication of Wolbachia in insect cell lines, artificial introduction of Wolbachia into novel hosts, and analysis of Wolbachia gene functions. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.
List of contents
Wolbachia: Advancing into a second century.- Investigation of cytoplasmic incompatibility patterns in Culex pipiens: from field samples to laboratory isofemale lines.- Maintenance and Evaluation of Wolbachia Male-Killers of Dipterans.- Examining Wolbachia-Induced Parthenogenesis in Hymenoptera.- Pillbug: a toolbox for feminisation.- Detection, Quantification and Elimination of Wolbachia in Bed Bugs.- Detection and analysis of Wolbachia in plant-parasitic nematodes and insights into Wolbachia evolution.- Developing Wolbachia-infected cell lines from insects.- Use of antibiotics to eliminate Wolbachia from mosquitos and cell culture.- Preparation of infectious Wolbachia from a mosquito cell line.- Wolbachia transinfection via embryonic microinjection.- Measuring host fitness effects and transmission of Wolbachia strains in Aedes aegypti mosquitoes.- Detection of natural Wolbachia strains in Anopheles mosquitoes.- Procedures for detection of Wolbachia-conferred antiviral protection in Drosophila melanogaster.- Validating a mitochondrial sweep accompanying the rapid spread of a maternally-inherited symbiont.- Screening host genomic data for Wolbachia infections.- A worked example of screening genomic material for the presence of Wolbachia infection.- Use of Drosophila Transgenics to Identify Functions for Symbiont Effectors.- Using Baker's Yeast to Determine Functions of Novel Wolbachia (and other prokaryotic) Effectors.- Isolation of Phage WO Particles from Wolbachia-infected Arthropods.- A light in the dark: Uncovering Wolbachia-host interactions using fluorescence imaging.
