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Colicin A hybrids: a genetic tool for selection of type II secretion‐proficient Pseudomonas strains
Author(s) -
Voulhoux Romé,
Lazdunski Andrée,
Filloux Alain
Publication year - 2001
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.1093/embo-reports/kve010
Subject(s) - periplasmic space , colicin , secretion , biology , bacterial outer membrane , signal peptide , type vi secretion system , gram negative bacteria , bacteria , heterologous , pseudomonas aeruginosa , gene , phenotype , secretory protein , type three secretion system , genetics , microbiology and biotechnology , plasmid , peptide sequence , mutant , biochemistry , escherichia coli , virulence
The Gram‐negative bacterium Pseudomonas aeruginosa secretes the majority of its extracellular proteins by the type II secretion mechanism, a two‐step process initiated by translocation of signal peptide‐bearing exoproteins across the inner membrane. The periplasmic forms are transferred across the outer membrane by a machinery consisting of 12 xcp gene products. Although the type II secretion machinery is conserved among Gram‐negative bacteria, interactions between the secreted proteins and the machinery are specific. The lack of a selectable phenotype has hampered the development of genetic strategies for studying type II secretion. We report a novel strategy to identify rare events, such as those that allow heterologous secretion or identification of extragenic suppressors correcting xcp defects. This is based on creating a host‐vector system where the non‐secretory phenotype is lethal. The original tool we designed is a hybrid protein containing elastase and the pore‐forming domain of colicin A.