Production of Ribosomal Protein S12/Renilla Luciferase Fusion and Development of a Bioluminescent Method for Detection of Aminoglycosides in Pork and Studying Its Recognition Mechanism

Abstract

In this study, the genes of Escherichia coli ribosomal protein S12 and renilla luciferase were linked and expressed to produce a fusion protein, and its intermolecular interactions and affinities with sevenaminoglycosides were studied. Then, the fusion protein was used as the core agent to develop a bioluminescent method on a conventional microplate for determination of the residues of thesevenaminoglycosides in pork. This method contained only one sample-loading step, and thus the assay was finished within 30 min. The limits of detection for the sevendrugs were in the range of 0.51–1.1 ng/mL, and the sensitivity for a specific drug was mainly determined by the receptordrug affinity but not related with the binding energy. After general comparison, the present method showed generally better performances than the previously reported enzyme-linked immunosorbent assays for aminoglycosides. This is the first study reporting the recognition mechanisms of Escherichia coli ribosomal protein S12 for aminoglycosides and developing a bioluminescent method for detection of aminoglycoside residues in pork samples.