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SummaryFrom studies of behaviour, chemical communication, genomics and developmental biology, among many others, honey bees have long been a key organism for fundamental breakthroughs in biology. With a genome sequence in hand, and much improved genetic tools, honey bees are now an even more appealing target for answering the major questions of evolutionary biology, population structure, and social organization.At the same time, agricultural incentives to understand how honey bees fall prey to disease, or evade and survive their many pests and pathogens, have pushed for a genetic understanding of individual and social immunity in this species. Below we describe and reference tools for using modern molecular-biology techniques to understand bee behaviour, health, and other aspects of their biology. We focus on DNA and RNA techniques, largely because techniques for assessing bee proteins are covered in detail in Hartfelder et al. (2013). We cover practical needs for bee sampling, transport, and storage, and then discuss a range of current techniques for genetic analysis. We then provide a roadmap for genomic resources and methods for studying bees, followed by specific statistical protocols for population genetics, quantitative genetics, and phylogenetics. Finally, we end with three important tools for predicting gene regulation and function in honey bees: Métodos estándar para la investigación molecular en Apis mellifera ResumenLas abejas de miel han sido durante mucho tiempo un organismo clave para avances fundamentales en biología a partir de estudios de su comportamiento, comunicación química, genómica y de biología del desarrollo, entre otros muchos. Con la secuencia del genoma en la mano y herramientas genéticas mucho mejores, las abejas son ahora un blanco aún más atractivo para responder a las preguntas más importantes de la biología evolutiva, la estructura de las poblaciones y la organización social. Al mismo tiempo, los incentivos agrícolas para entender cómo las abejas caen enfermas, o evadir y sobrevivir a sus muchas plagas y patógenos, han presionado para comprender genéticamente la inmunidad individual y social en esta especie. A continuación se describen y se hace referencia a herramientas que hacen uso de modernas técnicas de biología molecular para entender el comportamiento de las abejas, su salud y otros aspectos de su biología. Nos centramos en las técnicas de ADN y ARN, en gran parte debido a que las técnicas de evaluación de las proteínas de la abeja se tratan en detalle en Hartfelder et al. (2013). Cubrimos las necesidades prácticas de toma de muestras de abejas, su transporte y almacenamiento, y luego se discuten una serie de técnicas actuales de análisis genético. A continuación, se proporciona una hoja de ruta para los recursos genómicos y métodos para estudiar las abejas, seguido de protocolos estadísticos específicos de la genética de poblaciones, la genética cuantitativa y la filogenia.Finalmente, se termina con tres herramientas importantes para predecir la regulación génica y la fu...
The purple sea urchin, Strongylocentrotus purpuratus, possesses a sophisticated innate immune system that responds to microbes effectively by swift expression of the highly diverse Sp185/333 gene family. The Sp185/333 proteins are predicted to have anti-pathogen functions based on inducible gene expression and their significant sequence diversity. Sp185/333 proteins are all predicted to be intrinsically disordered and do not exhibit sequence similarities to other known proteins. To test the anti-pathogen hypothesis, a recombinant Sp185/333 protein, rSp0032, was evaluated and found to exhibit specific binding to marine Vibrio diazotrophicus and to Saccharomyces cerevisiae, but not to two Bacillus species. rSp0032 also binds to LPS, β-1,3-glucan and flagellin but not to peptidoglycan. rSp0032 binding to LPS can be competed by LPS, β-1,3-glucan and flagellin but not by peptidoglycan. We speculate that the predicted intrinsically disordered structure of rSp0032 may adapt to different conformations in binding to a limited number of PAMPs and pathogens. Given that rSp0032 binds to a range of targets, and that up to 260 different Sp185/333 proteins can be expressed per individual sea urchin, this family of immune response proteins may facilitate effective host protection against a broad array of potential pathogens encountered in the marine environment.
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