Dr. Andrea Castillo
Helicobacter pylori, a bacterial gastric pathogen infecting approximately 50% of the human population, produces gastritis, ulcers, and gastric cancers. Colonizing the inhospitable and fluctuating environment in the stomach requires tight genetic control. However, H. pylori lacks many genetic regulatory elements present in other bacteria to control gene expression. Instead, over 200 small RNAs (sRNAs; noncoding RNAs shorter than 300 nucleotides) have been found in this bacterium, but few have been fully characterized. Of those, many are antisense to virulence genes. Characterizing these sRNAs is important in understanding the mechanisms of molecular genetics and potentially supporting medical management of this pathogen. In the current study, a previously identified but as-yet uncharacterized sRNA was analyzed through reverse-transcription polymerase chain reaction (RT-PCR) utilizing primer walking, a technique employing custom oligonucleotides to experimentally determine the beginning and end regions of a transcript. RT-PCR results are unclear, representing longer-than-expected transcripts of variable lengths. This sRNA occurs downstream of another predicted sRNA. The results may represent two overlapping (possibly rather large) transcripts. Future work includes RT-PCR of two other antisense sRNAs and Northern blotting to analyze the size and gene boundaries of these sRNAs. Total results will explicate these three sRNAs and provide a foundation for further inquiry into the regulatory role these small but impactful molecules play in H. pylori.
McPeck, Roxanne and Castillo, Andrea, "Characterization of a Helicobacter pylori Small RNA by RT-PCR" (2022). 2022 Symposium. 11.
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