title: Functional cloning of Vibrio parahaemolyticus type III secretion system 1 in Escherichia coli K-12 strain as a molecular syringe.
abstract: The type III secretion system (T3SS) of gram-negative bacteria involves dedicated protein translocation machinery that directly injects proteins into target cells. Pathogenic bacteria already benefit from this unique system. The successful functional cloning of this useful tool into non-pathogenic bacteria would help establish novel clinical and basic biotechnology strategies in areas such as vaccine administration, the development of screening systems for anti-T3SS drugs and the target-specific delivery of bioactive compounds. In this study, we successfully cloned the Vibrio parahaemolyticus T3SS1 genetic locus into a non-pathogenic Escherichia coli K-12 strain. Assays performed here revealed that the T3SS1 cloned into the E. coli K-12 strain has the ability to translocate V. parahaemolyticus T3SS1 secreted proteins. Importantly, we also observed this system to allow the E. coli K-12 strain to inject foreign protein, as well as the V. parahaemolyticus T3SS effector, into cultured cells. These results demonstrate a prospective useful tool with experimental and therapeutic applications.
authors.affiliation: Laboratory of Clinical Research on Infectious Diseases, International Research Center for Infectious Diseases, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan. akeda@biken.osaka-u.ac.jp
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Akeda
authors.forename: Tomomi
authors.initials: T
authors.lastname: Kimura
authors.forename: Aiko
authors.initials: A
authors.lastname: Yamasaki
authors.forename: Toshio
authors.initials: T
authors.lastname: Kodama
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Iida
authors.forename: Takeshi
authors.initials: T
authors.lastname: Honda
authors.forename: Kazunori
authors.initials: K
authors.lastname: Oishi
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D058947:Bacterial Secretion Systems / Q000235:genetics*; D001428:Bacterial Vaccines / Q000008:administration & dosage; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D056426:Genetic Loci; D006367:HeLa Cells; D006801:Humans; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D014735:Vibrio Infections / Q000517:prevention & control; D014736:Vibrio parahaemolyticus / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems; D001428:Bacterial Vaccines; D011993:Recombinant Fusion Proteins