Construction of A Shuttled Plasmid Inducibly Expressing Green Fluorescent Protein and Its Application in Fluorescence Tracing
Fluorescence labeling of bacteria is an important and commonly used experimental technique,which is used to study the physiological and biochemical characteristics of bacteria and trace bacteria in infected hosts and cells.Bacterial fluorescent labeling is usually achieved by the expression of fluorescent protein in plasmid.However,the constructed plasmid cannot be used in common due to different species and characteristics of bacteria.In this study,we constructed a shuttled plasmid pBT-iEGFP that induced expression of enhanced green fluorescent protein by anhydrous tetracycline.The results showed that the pBT-iEGFP plasmid was successfully induced for expression of green fluorescent protein in avian pathogenic Escherichia coli,Salmonella typhimurium and Brucella melitensis.The plasmid was stably inherited in bacteria for 5 blind passages.Induced expression of EGFP in intracellular Brucella confirmed that the pBT-iEGFP could be used to trace live Brucella in the process of cell infection.In summary,the plasmid pBT-iEGFP constructed in the study could be used in a variety of scientific experiments,which provided a tool for the research of bacterial fluorescence tracing.
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