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1.
Appl Microbiol Biotechnol ; 99(2): 729-39, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25038929

RESUMO

The Bluetongue virus (BTV) VP7 protein represents an important group-specific antigen that can serve as a basis for diagnostic tests. Here, we report the generation of a novel BTV group-specific monoclonal antibody (Mab; herein named 4H7) that recognizes a conformational epitope in the VP7 protein. We used a phage-displayed peptide screen and site-directed mutagenesis to define the VP7 amino acid residues that most strongly contribute to the conformational epitope recognized by Mab 4H7. Amino acid residues at positions 175, 185, 186, and 278 of the BTV VP7 protein strongly contributed to Mab 4H7 binding. These key amino acid residues are conserved among all BTV serotypes, whereas related Orbiviruses possess at least one amino acid substitution at these positions. We developed a competitive enzyme-linked immunosorbent assay (c-ELISA) using Mab 4H7 and recombinant BTV VP7 protein to detect serum antibodies against this BTV group-specific VP7 epitope. The c-ELISA was used to screen 833 clinical samples collected from animals in three provinces of China. BTV seroprevalence in the three provinces ranged from 25.42 to 47.45 %. This work provides the foundation for a new diagnostic c-ELISA that can be further applied to BTV surveillance activities and informs our understanding of the structure of the BTV VP7 protein.


Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos Antivirais/imunologia , Vírus Bluetongue/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Animais , Anticorpos Monoclonais/sangue , Anticorpos Antivirais/sangue , China , Clonagem Molecular , Epitopos/sangue , Epitopos/imunologia , Cabras/virologia , Mutagênese Sítio-Dirigida , Conformação Proteica , Proteínas Recombinantes/sangue , Proteínas Recombinantes/imunologia , Reprodutibilidade dos Testes , Estudos Soroepidemiológicos , Ovinos/virologia , Proteínas do Core Viral/sangue , Proteínas do Core Viral/imunologia
2.
Ecol Evol ; 3(10): 3447-54, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24223281

RESUMO

Wheat (Triticum aestivum L.) is one of the most successful domesticated plant species in the world. The majority of wheat carries mutations in the Puroindoline genes that result in a hard kernel phenotype. An evolutionary explanation, or selective advantage, for the spread and persistence of these hard kernel mutations has yet to be established. Here, we demonstrate that the house mouse (Mus musculus L.) exerts a pronounced feeding preference for soft over hard kernels. When allele frequencies ranged from 0.5 to 0.009, mouse predation increased the hard allele frequency as much as 10-fold. Studies involving a single hard kernel mixed with ∼1000 soft kernels failed to recover the mutant kernel. Nevertheless, the study clearly demonstrates that the house mouse could have played a role in the evolution of wheat, and therefore the cultural trajectory of humankind.

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