A method for rapid screening of bacterial quorum sensing inhibitors using luminescence

Abstract

The invention provides a method for rapidly screening a bacterial quorum sensing inhibitor (QSI) by utilizing a luminescence method. The method is characterized by infecting escherichia coli ER2738 carrying pSB536 by utilizing the characteristic that a phage M13 does not lyse a host bacterium and can be secreted outside cells and then screening a bacterial clone which is blue under visible light and does not emit or emits weak biological fluorescent light under exciting light in a plate to which a C4-AHL molecule, tetracycline, ampicillin and X-gal / IPTG are added; then amplifying the phage, carrying out streak culture in the plate to which the substances are added, verifying the functions of the QSI by a method of inhibiting bioluminescence and finally obtaining specific QSI polypeptide sequences through sequencing. The method is simple in steps, and multiple QSI candidate polypeptides can be obtained only within three days to about one weak, so that the method has very obvious advantages.

Description

technical field [0001] The invention relates to genetic engineering and aptamer technology, and discloses a phage display technology for screening bacterial quorum sensing inhibitors. Specifically, it refers to a technique for screening bacterial quorum sensing inhibitors, which is also applicable to the screening of quorum sensing molecular inhibitors such as AI-1 and short-chain N-acyl homoserine lactones (AHLs) . The phage display technology involved in the present invention can be applied to any phage library with random libraries, such as phage antibody library, cyclic peptide, linear polypeptide, etc. The host bacteria involved are male F' bacteria that can be infected by phage libraries, such as ER2738, DH5α F', XL1-Blue, etc., and the vectors involved are vectors that can sense QS molecules to cause bacterial luminescence, such as pSB536, pJBA130, pREC- FF etc. It belongs to the field of prevention and control of pathogenic microorganisms. Background technique ...

AI Technical Summary

Problems solved by technology

The above four are the main sources of obtaining QSI at present, but there are some problems: the sample components of plants and other sources are complex, the extraction, identification and purification process of QSI in the early stage are cumbersome, and they are also limited by uncertain factors such as sample purity, temperature, and region. ;Artificial design synthesis and antibody screening not only require professional chemical synthesis and structural analysis background, but also take into account the complexity of the sample preparation process, professional equipment, intermediate product toxicity and contamination assessment, and a series of issues such as sample purity

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