Experimental Phase of “The Curious Case of OipA”

Hello and welcome to the wrap up of the Fall 2016 edition of “The Curious Case of OipA!” Since I was last on the SuRGe blog, I have completed phase 1 of my project, and have successfully created all of my mutant oipA strains of Helicobacter pylori. Now I am well into the experimental phase, and have some interesting preliminary results regarding the role of OipA in the pathogenicity of both cag pathogenicity island negative (less virulent) and positive (more virulent) strains of HP.

Recently there has been a (potentially) exciting development in this phase! As I was reviewing my data last week and thinking about the story of OipA and how it relates to the Cag Pathogenicity Island, I realized that I was missing a key element in this narrative. For some reason it had never occurred to me that if I wanted to figure out how OipA impacts stomach cell binding and virulence in both cagPAI positive and cagPAI negative strains, I would need to compare Helicobacter pylori (HP) cell expression of OipA both in the presence and absence of AGS (stomach) cells. So far, I have only looked at oipA transcription in my HP mutants that were not exposed to AGS cells. This, however, does not take into account any adaptations that may occur to alternatively transcribe the gene in the presence of AGS cells. So I designed an assay meant to accomplish this measurement, and I tried it out for the first time this weekend. I will not know the results of it until this week, and I already discovered many tweaks that need to be made, but I am excited to see how it turns out!

If you would like more info on my project, feel free to check out http://honorsfellows.blogs.wm.edu/author/dnhorridge/