A New Method for Extracting Outer Membrane Vesicles of Campylobacter jejuni
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Abstract:
Outer membrane vesicles (OMVs) of Campylobacter jejuni are globular vesicles secreted to the outside of cellsduring the growth process, whichmainly contain outer membrane proteins and some periplasmic substances, and play an important role in the survival, colonization, communication between bacteria and host cells, and pathogenic mechanisms. Therefore, the development of an efficient extraction method is the key to studying the biological function of OMVs. This study found that it is most appropriate to extract OMVs after culturing C. jejuni with MH medium for 15 h under microaerobic conditions. Under optimal conditions, the ultrafiltration concentration method was used to extract OMVs from the bacterial liquid, and the extract was further filtered using a 0.22 μm microporous filter to remove flagella and other impurities. In order to characterize the quality of the extracted OMVs, a transmission electron microscope was used. It was found that the extracted OMVs had a typical morphology (with their size ranging from 50 to 300 nm), and contained less impurities. The results of SDS-PAGE showed that OMVs contained a large amount of proteins (whose total content was 40.50 mg/mL). Finally, the outer membrane protein antibody was used to prove successfully that the main components of the extracted material were OMVs. In this study, an extraction system for extracting C. jejuni OMVs from liquid culture medium was successfully established. Various analytical methods have proved that its extraction efficiency is high and its quality is good. It is suitable for subsequent studies on the biological functions of C. jejuni OMVs.
