Discipline: Biological Sciences
Josue A. Rodriguez Ramos - University of Puerto Rico, Mayaguez
Co-Author(s): Carlos Rios Velazquez and Moises de Jesus Cruz, University of Puerto Rico, Mayaguez, PR
Bacteriophages can be utilized as an efficient way to introduce genetic material into host cells. This process, known as transduction, is used as a bio-prospect in order to perform genetic and biochemical analysis. Transduction is also very useful in Biotechnology, where it is commonly used to mass produce desired molecules. Rhodobacter sphaeroides is a Purple-Non Sulfurous Bacteria (PNSB) with a very versatile metabolism. It can use different carbon sources for energy and is capable of growing under different conditions: both aerobically and anaerobically as well as by photosynthesis. Our laboratory has isolated and characterized several Rhodobacter sphaeroides specific bacteriophages capable of performing lysis. However, to date, no transduction capability has been detected, thus, limiting the manipulation for biotechnological applications of the bacteria. The goal of this research is to identify genetic targets in R. sphaeroides that are involved in the susceptibility of infection by the phages and the integration of the phage into the R. sphaeroides genome. A series of R. sphaeroides 2.4.1 and 630 Transposon mutants were generated to be tested for susceptibility of infection by seven different bacteriophages. To date, a total of 23 R. sphaeroides 2.4.1 and 105 R. sphaeroides 630 transposon mutants have been successfully created. Of these mutants, 18 R. sphaeroides mutants (10 from 630 and 8 from 2.4.1) were screened by means of viral plaque assays. A bacterial lawn was made and 2µL aliquots of 5 different phage solutions were placed on the plate in search for a change in viral infection by the previously isolated R. sphaeroides specific bacteriophages. A change in infection pattern between the mutant and the wildtype would evidence that during mutagenesis, the transposon inactivated genes whose function mediates or influences this infection process, but no difference in phage activity was demonstrated between the R. sphaeroides transposon mutants and the R. sphaeroides wild type. Further viral plaque assays could reveal potential genetic targets involved in bacteriophage susceptibility.
Funder Acknowledgement(s): University of Puerto Rico, Mayaguez Department of Biology; Puerto Rico Louis Stokes Alliance for Minority Participation (PRLSAMP).
Faculty Advisor: Carlos Rios Velazquez, firstname.lastname@example.org
Role: As the primary researcher for this project, every aspect of it has been performed by myself. Everything from the experimental design to the data collection of this investigation has been of my own doing.