As loading control and control for cell lysis, the bacterial heat shock protein DnaK was detected. In Torin 1 chemical structure total cell lysates, we observed a non-specific binding (indicated by the asterisk). (DOC 30 KB) Additional file 2: Quantification of the effects of various deletions in sseB on synthesis and secretion of SseB in vitro and on secretion and partitioning of SseD in vitro. The signals of Western blot shown in Fig. 2 for the secretion and partitioning of SseB
and mutant variant and the Western blot shown in Fig. 3 for the effector of deletions in SseB on secretion an partitioning of SseD were quantified. Densitometry was performed using ImageJ software http://rsbweb.nih.gov/ij/ and signal intensities were normalized to the total cell fraction set to 100%. (TIFF 605 KB) Additional file 3: Oligonucleotides used in this study. The designation and sequence of oligonucleotides used for mutagenesis, strain construction and sequencing is shown. (DOC 33 KB) References 1. Gerlach RG, Hensel M: Protein secretion systems and adhesins: the molecular armory of Gram-negative pathogens. Int J Med Microbiol 2007,297(6):401–415.PubMedCrossRef 2. Galan JE, Wolf-Watz H: Protein delivery into eukaryotic cells by type III secretion machines. Nature 2006,444(7119):567–573.PubMedCrossRef
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