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We identified an open reading frame (ORF) in Burkholderia pseudomallei that resembles Staphylococcus aureus Biofilm-associated protein (Bap). We designated this B. pseudomallei ORF Bap-like protein (Blp). Biofilm-associated proteins are surface exposed molecules that are involved in adherence and biofilm formation. In order to study the function of B. pseudomallei Blp, we wanted to express the protein in a heterologous expression system. B. thailandensis is genetically similar to B. pseudomallei, but is relatively avirulent and does not possess the locus encoding Blp. A plasmid was created, designated pWGBtBlp, harboring blp and regions flanking the gene that may encode factors needed for secretion of Blp. B. thailandensis pWGBtBlp produces more biofilm and adheres to epithelial cells at higher levels than the vector control; however, a Tn insertion in the blp gene on the plasmid did not result in wild-type levels of biofilm production and adherence in B. thailandensis, suggesting Blp is not responsible for these processes. In order to determine the factor mediating these phenotypes, we generated plasmids containing portions of the blp locus and identified a small ORF with no known homologues that is able mediate biofilm production and adherence independently of any additional factors. |
