Verifying Expression of Helicobacter pylori sRNAs HPnc2450 and HPnc2620 Using Northern Blot Analysis
Faculty Mentor
Andrea Castillo, PhD
Presentation Type
Poster
Start Date
May 2025
End Date
May 2025
Location
PUB NCR
Primary Discipline of Presentation
Biology
Abstract
Helicobacter pylori is a bacterial pathogen that infects the stomachs of half the world’s population and has the potential to cause severe illness, including ulcers and cancer. Factors contributing to disease outcomes include genetic variability of the infecting H. pylori strains and their regulation of gene expression. There is mounting evidence that small RNA (sRNA) molecules contribute to gene expression regulation in H. pylori. A former graduate student contributed to this body of work with the characterization of sRNAs HPnc2540 and HPnc2620. She compared by RNA sequencing the RNA from an unmodified H. pylori strain to RNA of those overexpressing HPnc2540 or HPnc2620, and found that these sRNAs regulate expression of a set of overlapping genes, including some that promote disease. Our objective is to use Northern Blotting to demonstrate that HPnc2540 and HPnc2620 expression is higher in the respective H. pylori overexpression strains compared to the unmodified strain. This involves gel electrophoresis on RNA isolated from three strains of H. pylori: an unmodified strain, a HPnc2540-overexpression strain, and a HPnc2620-overexpression strain. The gels will then be blotted to a nylon membrane and “probed” with a fluorescent oligonucleotide that is complementary to HPnc2540 or HPnc2620. The probed nylon membranes will be imaged by a Typhoon NIH Biomolecular Imager and the intensity of fluorescent bands will indicate the level of expression of the sRNA in each strain. We anticipate that fluorescent bands will be brighter in H. pylori sRNA-overexpression strains probed with the complementary fluorescent oligonucleotide than in the unmodified strain.
Recommended Citation
Potts, Kris V. and Castillo, Andrea PhD, "Verifying Expression of Helicobacter pylori sRNAs HPnc2450 and HPnc2620 Using Northern Blot Analysis" (2025). 2025 Symposium. 16.
https://dc.ewu.edu/srcw_2025/ps_2025/p1_2025/16
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Verifying Expression of Helicobacter pylori sRNAs HPnc2450 and HPnc2620 Using Northern Blot Analysis
PUB NCR
Helicobacter pylori is a bacterial pathogen that infects the stomachs of half the world’s population and has the potential to cause severe illness, including ulcers and cancer. Factors contributing to disease outcomes include genetic variability of the infecting H. pylori strains and their regulation of gene expression. There is mounting evidence that small RNA (sRNA) molecules contribute to gene expression regulation in H. pylori. A former graduate student contributed to this body of work with the characterization of sRNAs HPnc2540 and HPnc2620. She compared by RNA sequencing the RNA from an unmodified H. pylori strain to RNA of those overexpressing HPnc2540 or HPnc2620, and found that these sRNAs regulate expression of a set of overlapping genes, including some that promote disease. Our objective is to use Northern Blotting to demonstrate that HPnc2540 and HPnc2620 expression is higher in the respective H. pylori overexpression strains compared to the unmodified strain. This involves gel electrophoresis on RNA isolated from three strains of H. pylori: an unmodified strain, a HPnc2540-overexpression strain, and a HPnc2620-overexpression strain. The gels will then be blotted to a nylon membrane and “probed” with a fluorescent oligonucleotide that is complementary to HPnc2540 or HPnc2620. The probed nylon membranes will be imaged by a Typhoon NIH Biomolecular Imager and the intensity of fluorescent bands will indicate the level of expression of the sRNA in each strain. We anticipate that fluorescent bands will be brighter in H. pylori sRNA-overexpression strains probed with the complementary fluorescent oligonucleotide than in the unmodified strain.
