Process Overview
Despite our results attempting to propagate the phage, the experimental process below outlines our projected methods. After propagating the phi EA104 bacteriophage, we would have used the following methods:
Steps
In Silica Work
- We used the annotated phi EA 104 phage under the Kolesnikvirus taxonomic family and place it into Benchling after downloading from NCBI.
- Next, we deisgned a gene cassette focused on cloning the phage as an insert into a BAC/YAC capture vector in order to grow the plasmid in E.coli. Our end goal is to cut out a gene essential for the maturation of the phage genome, yet will still allow already developed phages to infect our target bacteria. This aims to make it so that only phages propagated in a lab setting can fully develop into a mature form. This biocontains the phages that have been released into the environment to ensure that they do not replicate and overtake other microbes once the target bacteria
(E.amylovora) has been eradicated. - Our gene cassette used guide RNAs and arms of homology (hooks) to use the power of homologous recombination with the vector.
Conclusion
Summarize the process and discuss any final thoughts.