Oropouche virus (OROV) is the unique known human pathogen belonging to serogroup Simbu of Orthobunyavirus genus and Bunyaviridae family. OROV is transmitted by wild mosquitoes species to sloths, rodents, monkeys and birds in sylvatic environment, and by midges (Culicoides paraensis and Culex quinquefasciatus) to man causing explosive outbreaks in urban locations. OROV infection causes dengue fever-like symptoms and in few cases, can cause clinical symptoms of aseptic meningitis. OROV contains a tripartite negative RNA genome encapsidated by the viral nucleocapsid protein (NP), which is essential for viral genome encapsidation, transcription and replication. Here, we reported the first study on the structural properties of a recombinant NP from human pathogen Oropouche virus (OROV-rNP). OROV-rNP was successfully expressed in E. coli in soluble form and purified using affinity and size-exclusion chromatographies. Purified OROV-rNP was analyzed using a series of biophysical tools and molecular modeling. The results showed that OROV-rNP formed stable oligomers in solution coupled with endogenous E. coli nucleic acids (RNA) of different sizes. Finally, electron microscopy revealed a total of eleven OROV-rNP oligomer classes with tetramers (42%) and pentamers (43%) the two main populations and minor amounts of other bigger oligomeric states, such as hexamers, heptamers or octamers. The different RNA sizes and nucleotide composition may explain the diversity of oligomer classes observed. Besides, structural differences among bunyaviruses NP can be used to help in the development of tools for specific diagnosis and epidemiological studies of this group of viruses.
RESUMEN Theobroma cacao L. es la única especie del género Theobroma que se explota comercialmente en grandes extensiones, registrando en la actualidad una amplia distribución mundial, a través de programas de desarrollo directamente influenciados por factores vinculados al mercado y por los intereses de productores, comerciantes, industriales y consumidores. En este estudio se evaluaron 12 clones de cacao por medio de marcadores moleculares utilizando 10 secuencias microsatélites (SSRs), como un estudio piloto antes de un ensayo a mayor escala. Los datos se procesaron mediante el programa Power Marker Versión 3.25. Se estimaron las frecuencias alélicas y luego se generó una matriz de distancias genéticas basada en el coeficiente de Nei. Utilizando el algoritmo de agrupamiento UPGMA se generó el dendrograma correspondiente. El análisis de diversidad mostró un índice de diversidad genética total de 0,6944 considerado intermedio para los materiales evaluados. La estimación de la heterocigosidad promedio fue de 0,58579 y de promedio del contenido de información polimórfica de 0,6523. En este estudio los marcadores mTcCIR6, mTcCIR25, mTcCIR26 y mTcCIR12 son los más informativos y polimórficos. Se recomienda relacionar los resultados de diversidad genética con caracteres morfo-agronómicos y de patogenicidad en los distintos clones de cacao a fin de consolidar estrategias eficaces de mejoramiento y control de enfermedades.
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