Extraction of Bacterial Membrane Vesicle and Phage Complex by Density Gradient Ultracentrifugation
Shangru Li,
Anmin Ren,
Menglu Li,
Guobao Li,
Liang Yang,
Tianyuan Jia
Affiliations
Shangru Li
Shenzhen National Clinical Research Center for Infectious Disease, Shenzhen Third People’s Hospital, The Second Affiliated Hospital of Southern University of Science and Technology, Shenzhen, ChinaSchool of Medicine, Southern University of Science and Technology, Shenzhen, China
Anmin Ren
School of Medicine, Southern University of Science and Technology, Shenzhen, China
Menglu Li
Shenzhen National Clinical Research Center for Infectious Disease, Shenzhen Third People’s Hospital, The Second Affiliated Hospital of Southern University of Science and Technology, Shenzhen, ChinaSchool of Medicine, Southern University of Science and Technology, Shenzhen, China
Guobao Li
Shenzhen National Clinical Research Center for Infectious Disease, Shenzhen Third People’s Hospital, The Second Affiliated Hospital of Southern University of Science and Technology, Shenzhen, ChinaSchool of Medicine, Southern University of Science and Technology, Shenzhen, China
Liang Yang
Shenzhen National Clinical Research Center for Infectious Disease, Shenzhen Third People’s Hospital, The Second Affiliated Hospital of Southern University of Science and Technology, Shenzhen, ChinaSchool of Medicine, Southern University of Science and Technology, Shenzhen, China
Tianyuan Jia
Shenzhen National Clinical Research Center for Infectious Disease, Shenzhen Third People’s Hospital, The Second Affiliated Hospital of Southern University of Science and Technology, Shenzhen, ChinaSchool of Medicine, Southern University of Science and Technology, Shenzhen, China
The bacterial membrane vesicles (MVs) are non-replicative, nanoscale structures that carry specific cargos and play multiple roles in microbe–host interactions. An appropriate MV isolation method that mimics complex pathogen infections in vivo is needed. After bacterial MVs extraction, flagella or pili can be frequently observed along with MVs by transmission electron microscope (TEM). Recently, MVs from Pseudomonas aeruginosa were found to coexist with Pf4 phages, and this MV–phages complex exhibited a different impact on host cell innate immunity compared with MVs or phages solely. The presence of this MVs–phages complex simulates the real condition of complex pathogen infections within the host. This protocol outlines the extraction of the MVs and Pf4 phages complex of P. aeruginosa PAO1, including the respective isolation and qualification approaches. Our step-by-step bacterial MVs–phages complex extraction protocol provides valuable insights for further studying microbe–host cell interactions and the development of novel phage therapies.
