Results

Overview


Portal proteins exist within the Erwinia phage genome with portal complexes that have different contortions per each stage of phage assembly (Prevelige, 1993). This potential target protein is key in capsid assembly and the DNA packing process (Dedeo, 2019). Potential infection and bacteriophage development catalyzation are directly corresponded to these portal proteins. Further experimental research and redesign would allow for the exploration of deleting this gene from the bacteriophage genome to regulate the progression of infection. Another potential direction that could be further explored includes induction of anaerobic respiration to potentially reduce the ability of E. amylovora to survive. E. amylovora have genes involved in anaerobic respiration, and during these metabolic processes, the genome size can become reduced or experience loss and decomposition of genes responsible for growth cycles (Sebaihia, 2010). To further test and examine these phenotypic-genotypic relationships, future directions include using conditions of limited oxygen supply cultured in a hypersaline environment to describe bacteriophage progression when in anaerobic conditions (Muller, 2005).


Although this project was unable to successfully eliminate the phiEa104 bacteriophage by way of Erwinia amylovora bacteriophage genomic extraction, this investigation elucidated potential new target sites, replacements for pesticides, and ideas for biocontainment tactics against microbial attacks. As mentioned above, our team has explored other potential avenues for targeting genomic deletion to mitigate bacteriophage assembly. Hypothetical implications corroborate the necessity of development within fields of containing bacterial infection among crops. Furthermore, based on our formulative research, targeting proteins that control the morphology of the virus could mediate growth phases and assembly of viral infection. Ultimately, cloning techniques, genomic extraction, and the power of homologous recombination have the potential to eliminate Erwinia amylovora as a threat entirely.

Project Achievements


  • Inoculated bacterial liquid culture with virus for genomic preparation.
  • Effectively carried out viral infection of liquid cultures to generate viral lysate for genomic extraction.
  • Successfully produced viral plates containing bacterial cultures that had phage plaques.
  • Identified viral plaque formation on plates.
  • Able to obtain DNA via genomic extraction of phage after outgrowth.

Unsuccessful Segments


Ultimately, the Erwinia amylovora bacteria and specified phage do not interact to a desirable level for utilization as hypothesized.

  • Bacteria and phage not compatible by way of current experimental design methods in terms of safety protocols. Next steps would be to use phenylchloroform, which is a highly dangerous carcinogen.
  • Projected targets for genomic extraction from DNA not reached due to time constraints after trying various methods to get reasonable DNA concentrations to continue experimentation.

Sources