Open AccessRestoring virulence to mutants lacking subunits of multiprotein machines: functional complementation of a Brucella virB5 mutant
Author(s) -
Sprynski Nicolas,
Felix Christine,
O’Callaghan David,
Vergunst Annette C.
Publication year - 2012
Publication title -
febs open bio
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.718
H-Index - 31
ISSN - 2211-5463
DOI - 10.1016/j.fob.2012.03.003
Subject(s) - complementation , mutant , virulence , replicon , plasmid , biology , genetics , expression vector , protein fragment complementation assay , multiprotein complex , gene , recombinant dna
Complementation for virulence of a non‐polar virB5 mutant in Brucella suis 1330 was not possible using a pBBR‐based plasmid but was with low copy vector pGL10. Presence of the pBBR‐based replicon in wildtype B. suis had a dominant negative effect, leading to complete attenuation in J774 macrophages. This was due to pleiotropic effects on VirB protein expression due to multiple copies of the virB promoter region and over expression of VirB5. Functional complementation of mutants in individual components of multiprotein complexes such as bacterial secretion systems, are often problematic; this study highlights the importance of using a low copy vector.