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1.
STAR Protoc ; 4(3): 102504, 2023 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-37585296

RESUMO

tRNA-bound amino acids often need to be identified, for instance, in cases where different amino acids compete for binding to the same tRNA. Here, we present a mass-spectrometry-based protocol to determine the amino acids bound to tRNA by aminoacylation. We detail how to perform the aminoacylation reaction, the preparation of the aminoacyl-tRNA for measurement, and the mass spectrometric analysis. We use arginine acylation as an example; however, this protocol can be applied to any other amino acid.


Assuntos
Aminoácidos , Aminoacil-tRNA Sintetases , Aminoácidos/metabolismo , Aminoacilação , Aminoacil-tRNA Sintetases/química , RNA de Transferência/metabolismo , Espectrometria de Massas
2.
PLoS One ; 10(5): e0124824, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25954946

RESUMO

The aerodynamic features of a bio-realistic 3D fruit fly wing in steady state (snapshot) flight conditions were analyzed numerically. The wing geometry was created from high resolution micro-computed tomography (micro-CT) of the fruit fly Drosophila virilis. Computational fluid dynamics (CFD) analyses of the wing were conducted at ultra-low Reynolds numbers ranging from 71 to 200, and at angles of attack ranging from -10° to +30°. It was found that in the 3D bio-realistic model, the corrugations of the wing created localized circulation regions in the flow field, most notably at higher angles of attack near the wing tip. Analyses of a simplified flat wing geometry showed higher lift to drag performance values for any given angle of attack at these Reynolds numbers, though very similar performance is noted at -10°. Results have indicated that the simplified flat wing can successfully be used to approximate high-level properties such as aerodynamic coefficients and overall performance trends as well as large flow-field structures. However, local pressure peaks and near-wing flow features induced by the corrugations are unable to be replicated by the simple wing. We therefore recommend that accurate 3D bio-realistic geometries be used when modelling insect wings where such information is useful.


Assuntos
Simulação por Computador , Drosophila/fisiologia , Voo Animal/fisiologia , Asas de Animais/diagnóstico por imagem , Microtomografia por Raio-X , Animais , Processamento de Imagem Assistida por Computador , Análise Numérica Assistida por Computador , Pressão , Reprodutibilidade dos Testes
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