Using a Mutagenesis Strategy to Identify Dissimilatory Sulfate Reducing Genes in the Gut Bacterium Desulfovibrio piger
Faculty Mentor
Andrea R Castillo
Presentation Type
Poster
Start Date
May 2025
End Date
May 2025
Location
PUB NCR
Primary Discipline of Presentation
Biology
Abstract
Desulfovibrio piger is an anaerobic, sulfate-reducing bacterium naturally found in the human colon. It produces hydrogen sulfide (H2S) as a byproduct of its dissimilatory sulfate reduction (DSR) metabolism, playing a role in gut health and immune function. Excessive growth of D. piger is correlated with excess H2S levels, which damages gut epithelial cells and may contribute to inflammatory diseases such as multiple sclerosis (MS) and inflammatory bowel disease. Specific genes involved in sulfate reduction and H2S synthesis in D. piger remain largely unidentified. To address this gap, our goal is to develop UV mutagenesis and H2S detection protocols for D. piger, followed by genome sequencing of DNA from H2S defective strains, that will allow us to identify genes essential for DSR. We are developing this protocol for the challenging-to-culture D. piger by first practicing experimental techniques with the model bacterium E. coli. Thus far, serially diluted E. coli has been irradiated in several independent kill curve experiments that correlate UV exposure with percent bacterial killing. After some troubleshooting with the UV machine, we are poised to conduct these experiments with D. piger. Mutagenized D. piger cells will then be cultured on Thiosulfate Citrate Bile Salts Sucrose (TCBS) agar to identify the ones defective in H2S production; H2S producers form black colonies and H2S mutants form white colonies. Genome sequencing of DNA from white colonies will identify the mutated genes, and thus those involved in H2S production.
Recommended Citation
Simpson, Justin M.; Ziegler, Zac E.; and Castillo, Andrea R., "Using a Mutagenesis Strategy to Identify Dissimilatory Sulfate Reducing Genes in the Gut Bacterium Desulfovibrio piger" (2025). 2025 Symposium. 19.
https://dc.ewu.edu/srcw_2025/ps_2025/p2_2025/19
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Using a Mutagenesis Strategy to Identify Dissimilatory Sulfate Reducing Genes in the Gut Bacterium Desulfovibrio piger
PUB NCR
Desulfovibrio piger is an anaerobic, sulfate-reducing bacterium naturally found in the human colon. It produces hydrogen sulfide (H2S) as a byproduct of its dissimilatory sulfate reduction (DSR) metabolism, playing a role in gut health and immune function. Excessive growth of D. piger is correlated with excess H2S levels, which damages gut epithelial cells and may contribute to inflammatory diseases such as multiple sclerosis (MS) and inflammatory bowel disease. Specific genes involved in sulfate reduction and H2S synthesis in D. piger remain largely unidentified. To address this gap, our goal is to develop UV mutagenesis and H2S detection protocols for D. piger, followed by genome sequencing of DNA from H2S defective strains, that will allow us to identify genes essential for DSR. We are developing this protocol for the challenging-to-culture D. piger by first practicing experimental techniques with the model bacterium E. coli. Thus far, serially diluted E. coli has been irradiated in several independent kill curve experiments that correlate UV exposure with percent bacterial killing. After some troubleshooting with the UV machine, we are poised to conduct these experiments with D. piger. Mutagenized D. piger cells will then be cultured on Thiosulfate Citrate Bile Salts Sucrose (TCBS) agar to identify the ones defective in H2S production; H2S producers form black colonies and H2S mutants form white colonies. Genome sequencing of DNA from white colonies will identify the mutated genes, and thus those involved in H2S production.
