Journal: Gut Pathogens
Article Title: Overexpression of serine protease HtrA enhances disruption of adherens junctions, paracellular transmigration and type IV secretion of CagA by Helicobacter pylori
Figure Lengend Snippet: Chromosomal introduction of a second, inducible htrA gene copy in H. pylori. a Schematic presentation of the IPTG-inducible htrA 26695 expression construct with the chloramphenicol acetyl-transferase (CAT) cassette ( top ), which was cloned into the plasticity region of H. pylori between genes HP0999 and HP1000 ( bottom ). b This construct was transformed into H. pylori wild-type ( Hp wt) strains 26695 and P12, and the expression of HtrA was investigated after 24 h growth in BHI medium in the presence or absence of 1 or 2 mM IPTG, respectively. The results for strain 26695 are shown. Western blotting using α-FlaA and α-CagA antibodies served as controls, indicating that equal amounts of proteins are present in the samples. c The band intensities of HtrA proteins were quantified densitometrically. The relative protein expression is given in “fold change”
Article Snippet: Antibodies The following antibodies were purchased: rabbit polyclonal α-CagA antibody (Austral Biologicals, San Ramon, CA/USA), monoclonal pan-phosphotyrosine α-PY99 (Santa Cruz, Santa Cruz, CA/USA), rabbit α-GAPDH (Santa Cruz), rabbit α-H. pylori (Dako, Glostrup/Denmark) and two monoclonal antibodies directed against the extracellular domain of E-cadherin, H-108 (Santa Cruz) and CD324 (BD Biosciences, San Jose, CA/USA).
Techniques: Expressing, Construct, Chloramphenicol Acetyltransferase Assay, Clone Assay, Transformation Assay, Western Blot