RESUMO
We developed and validated a novel lateral flow assay for the detection of crustacean protein in processed foods. This assay had high sensitivity; the visual detection limit for shrimp protein extract was 25µg/L, equivalent to 1µg/g protein in a food sample, and results could be obtained within 20min without sophisticated procedures or expensive equipment. Concordance between our assay and another validated quantitative enzyme-linked immunosorbent assay was 97% for commercially processed foods. This assay is rapid, simple, reliable, and highly correlated with validated enzyme-linked immunosorbent assays and is thus suitable for monitoring of food products, especially in food-processing facilities.
Assuntos
Proteínas de Artrópodes/análise , Crustáceos/química , Contaminação de Alimentos/análise , Imunoensaio/métodos , Alérgenos/análise , Alérgenos/imunologia , Animais , Proteínas de Artrópodes/imunologia , Crustáceos/imunologia , Fast Foods/análise , Imunoensaio/instrumentação , Limite de DetecçãoAssuntos
Genes Bacterianos/genética , Sequências Reguladoras de Ácido Ribonucleico/genética , Subunidades Ribossômicas/química , Subunidades Ribossômicas/metabolismo , Sequência de Bases , Códon de Iniciação/genética , Cinética , Dados de Sequência Molecular , RNA Bacteriano/genética , RNA Bacteriano/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA de Transferência/genética , RNA de Transferência/metabolismo , Subunidades Ribossômicas Maiores de Bactérias/metabolismo , Subunidades Ribossômicas Menores de Bactérias/metabolismoRESUMO
Translation initiation is the most dynamic and important step along a series of protein synthesis processes. In bacteria, it is generally accepted that the 70S ribosome initially dissociates into the 30S and 50S subunit, and then, the 30S ribosomal subunit binds to the Shine-Dargalno (SD) sequence of mRNA. We analyzed binding kinetics of 70S, 50S and 30S ribosomes to the SD sequence by using a mRNA-immobilized 27 MHz quartz-crystal microbalance (QCM). The 70S ribosome was found to bind strongly to the SD sequence as a ratio of 1:1 without dissociation to each subunit from the lateral side, as well as the 30S subunit. The binding constant for 70S increased in the presence of the initiator tRNA, which suggests that the SD and initiator codon of AUG could be also recognized precisely with 70S.
