Construction of in vitro reconstitution system for inverse autotransporter Intimin / 中华微生物学和免疫学杂志
Chinese Journal of Microbiology and Immunology
;
(12): 191-195, 2023.
Artículo
en Chino
| WPRIM
| ID: wpr-995273
ABSTRACT
Objective:
To construct an in vitro reconstitution system for inverse autotransporters in order to further investigate their translocation mechanism.Methods:
Intimin from Escherichi coli was used as a model substrate. Spheroplasts were prepared from Escherichi coli strains overexpressing Intimin to induce the expression of Intimin. Recombinant β-barrel assembly machinery (BAM) complex was obtained and purified, and then proteoliposomes containing BAM were prepared. Following the digestion with proteinase K, the translocation was detected by SDS-PAGE.Results:
Spheroplasts were induced to express Intimin, and then BAM-containing proteoliposomes were added to the system. Compared with control and liposomes groups, the experimental group showed that Intimin was resistant to proteinase K treatment, indicating that Intimin was successfully translocated.Conclusions:
The translocation of Intimin required the participation of BAM complex. An in vitro reconstitution system for inverse autotransporters was constructed in this study, providing a method to study the translocation mechanism of inverse autotransporters.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Idioma:
Chino
Revista:
Chinese Journal of Microbiology and Immunology
Año:
2023
Tipo del documento:
Artículo
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