Method for rapidly screening bacterial quorum sensing inhibitor (QSI) by utilizing suicide gene

Abstract

The invention provides a method for rapidly screening a bacterial quorum sensing inhibitor (QSI) by utilizing a suicide gene. The method is characterized in that under natural light, bacteria not infected with a phage do not turn blue and bacteria infected with the phage turn blue; induced by an added lethal condition inducer, a specific polypeptide with the function of inhibiting QS does not start expression of the suicide gene because of inhibiting the activities of QS molecules, so the bacteria survive; a non-specific polypeptide can not inhibit the QS molecules, so the suicide gene starts lethal; then survival candidate clones are picked out, functional verification is further carried out and finally a DNA (deoxyribonucleic acid) sequence corresponding to a specific polypeptide sequence is obtained by a method of DNA sequencing, thus obtaining an efficient and specific QSI polypeptide. 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, and the method is also applicable to the screening of long and short chain N-acyl homoserine lactones (AHLs) quorum sensing molecular inhibitors. 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 a phage library, such as ER2738, DH5αF', XL1-Blue, and the vector involved is the vector pSB537, which can cause the death of bacteria induced by its own gene by adding QS molecules. It belongs to the field of prevention and control of pathogenic microorganisms. Background technique [0002] The sit...

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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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