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Phage Genome Evolution: Research on Bacterial Infection

Explore the biodegradation of pollutants and study phages infecting bacterial cells to advance environmental research. Investigate the genetic homology of phages and bacterial hosts for potential phage therapy development.

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Phage Genome Evolution: Research on Bacterial Infection

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  1. E3 Summer Project“Exploring Research in Engineering and Science” Presentor: Jesus A. Hernandez Gregory-Portland High School Dr. Kung-Hui Chu Assistant Professor, Department of Civil Engineering

  2. The TEAM • Working along side Armando Vital, Brownsville, TX • Working under Dr. Kung-Hui Chu, Environmental Engineering/Civil Engineering • Working in Dr. Ry Young’s Lab, Center for Phage Technology (CPT) under direction of Dr. Jason Gill • Working with assistance from Myunghee Kim and Do Gyun Lee, both PhD. Students in Dr. Chu’s Lab

  3. Dr. Kung-Hui Chu • Biodegradation and bioremediation of priority pollutants and emerging contaminants • Molecular quantification of microbial risk in water • Optimization of bioenergy production • Application of bioretention for stormwater runoff management • Advancing knowledge on microbial ecology of nitrogen and carbon cycles

  4. The Laboratory Work • Techniques used in molecular biology for working with the phages • This is a collaborative effort in the disciplines of applied microbiology, virology, and environmental engineering

  5. Research Question A tale of two phages: Does high degree of phage gene homology exist? Significance: Phage Genome Evolution

  6. Background on Phages • Phages or bacteriophages are viruses that infect bacterial cells. They are found everywhere. • Phages, like any other virus, cannot reproduce on their own. • The phage hijacks the cell’s machinery to reproduce progeny. • Phages are specific for their host bacteria.

  7. Lysis via a 3 component system in Mycobacteria • Aholin protein opens a pore in the cytoplasmic membrane • Thecreation of the pore triggers the release of endolysin • An esterase enzyme is also released that degrades the outer mycolic acid cell wall layer • YouTube - T4 Virus infecting a bacteria.

  8. What do we know about these two phages? • A phage , Phage RopaN4, was isolated in Germany 20 years ago • Threephages (Ropa1, Ropa2, Ropa3) were recently isolated from Chu’s laboratory • To our surprise, we found that 2 of 3 of these phages have high similarity in their DNA sequences to N4.99.99% Identical!!!  The other was very closely related.

  9. Discovery => If this is true, it would be a new finding in “Phage Genome Evolution” Approach:new personnel to repeat our work at other laboratory in order to rule out any cross contamination during isolation phages in Dr. Chu’s laboratory. 

  10. Primary Objective • Isolate and characterize phages that infect Rhodococcusopacus. • Collect activated sludge and soil samples that may contain phage. • Secondary Objective: • Collect some DNA

  11. Rhodococcusopacus • Rhodococcusopacusis a specific bacteria • Belongs to the family Actinomycetes, related to Mycobacterium • R. opacusis a rod, nonmotile, mycobacterium • This bacteria was used as the host to isolate phage in Chu’s laboratory

  12. Relevance/Impact of the Research • Potentially open a new research direction in phage genomics. • Some Rhodococcus species are pathogens. A better understanding of phages specific to Rhodococcus can enhance the development of phage therapy. • Phage treatment might be possible to minimize common sludge bulking problem in treatment plants.

  13. The Challenge for Science • The cell wall of the bacteria is difficult to disrupt. • First step is to see which phages can actually infect R. opacus • Rhodococcus-specific phages can lyse the cell.

  14. Research Work • Collecting soil samples • Learn aseptic techniques • Preparing media (broth/food) • Enriching samples for phage • Growing liquid cultures of bacteria • Plating out phage • Collecting plaques of phage on petri dishes

  15. Flow Chart of Lab Work

  16. Data to be Generated • Data that will help characterize the phages from our collection sites. • Comparing the characteristics of new phages to the ones currently available. • Finding phages that will successfully infect R. opacus

  17. Future Implications • Does the high similarity of phage sequences unique to Rhodococcusspecies? • Does the high similarity of phage sequences present concern G+ pathogens? • What are the implications of high conservation of phage sequences?

  18. Summary • Phages infect bacterial cells • Focus on phage that infect Rhodococcusopacus • Isolate and characterize phages • Further exploration

  19. Classroom Lesson/Activities • Physics • Chemistry & Biology

  20. Acknowledgements • TAMU E3 Program • National Science Foundation (NSF) • Nuclear Power Institute (NPI) • Texas Workforce Comission • Dr. Kung-Hui Chu and her lab • Dr. Ry Young and CPT • Dr. Jason Gill • Myunghee Kim and Do Gyun Lee • Armando Vital (partner)

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