Thank you! To elaborate on our future directions, our phage may be plated on bacterial lawns of bacteria species other than M. smegmatis. As far as the DNA goes, a Polymerase Chain Reaction, or PCR, test would give more insight in further narrowing down the cluster organization. To do this, we would use primers to compare BonaeVitae and Imvubu to our phage. One of the lanes would consist of EE cluster phage DNA and the ClaI restriction enzyme, while another lane would have B cluster phage DNA and ClaI restriction enzyme.
Maybe I missed it, but what was the reason you chose investigating the EE cluster specifically and it’s potential bacteriophages rather than some other bacteriophage type.
We chose the EE cluster based on the restriction enzyme tool’s suggestions from comparing restriction digest results with a database of known phages. The highest similarity was with BonaeVitae, an EE cluster phage. B and L3 cluster phages were also found to matches from the online tool, which is why we chose to analyse those as well.
I was a little confused when you talked about the fact that it was clear from the figure that it was cut into 8 parts by one of the restriction enzymes. Could you explain that to me a bit more?
Of course! ClaI is a restriction enzyme that cut our phage DNA. The phage DNA was shown in the DNA ladder column. In figure 2, in the ClaI column, there were eight clear bands, which represent cuts.
Plating KingNeptune onto different lawns of bacterial species would give us insight into our phage’s host range, or the specific species which a phage can succesfully infect. Due to the physical structures of bacteria and phages, there are a limited number of bacterial species that are viable for infection by each phage species. Their structures must be compatible to enable infection. We can visualize these structures through EM photographs, and learning the host range of a phage adds to knowledge about the relationship between bacteria and phages. In phage therapy, the host range is important to know because one of the main benefits of this treatment as opposed to antibiotics is the ability to target specific bacterial species rather than killing all bacteria in the body. This is good because some bacteria is healthy for humans to have, so being able to target only the illness-causing bacteria is less destructive to the health of our bodies. Knowing the host range of a phage means that it can be used to target those bacteria species in medicine.
I really liked your presentation! I was wondering, what further experiments can be done in terms of studying the DNA of the phage?
Thank you! To elaborate on our future directions, our phage may be plated on bacterial lawns of bacteria species other than M. smegmatis. As far as the DNA goes, a Polymerase Chain Reaction, or PCR, test would give more insight in further narrowing down the cluster organization. To do this, we would use primers to compare BonaeVitae and Imvubu to our phage. One of the lanes would consist of EE cluster phage DNA and the ClaI restriction enzyme, while another lane would have B cluster phage DNA and ClaI restriction enzyme.
*KingNeptune DNA would also be included in a lane
Maybe I missed it, but what was the reason you chose investigating the EE cluster specifically and it’s potential bacteriophages rather than some other bacteriophage type.
We chose the EE cluster based on the restriction enzyme tool’s suggestions from comparing restriction digest results with a database of known phages. The highest similarity was with BonaeVitae, an EE cluster phage. B and L3 cluster phages were also found to matches from the online tool, which is why we chose to analyse those as well.
I was a little confused when you talked about the fact that it was clear from the figure that it was cut into 8 parts by one of the restriction enzymes. Could you explain that to me a bit more?
Of course! ClaI is a restriction enzyme that cut our phage DNA. The phage DNA was shown in the DNA ladder column. In figure 2, in the ClaI column, there were eight clear bands, which represent cuts.
By plating them on different species, what would that tell you about the phage and its characteristics?
Plating KingNeptune onto different lawns of bacterial species would give us insight into our phage’s host range, or the specific species which a phage can succesfully infect. Due to the physical structures of bacteria and phages, there are a limited number of bacterial species that are viable for infection by each phage species. Their structures must be compatible to enable infection. We can visualize these structures through EM photographs, and learning the host range of a phage adds to knowledge about the relationship between bacteria and phages. In phage therapy, the host range is important to know because one of the main benefits of this treatment as opposed to antibiotics is the ability to target specific bacterial species rather than killing all bacteria in the body. This is good because some bacteria is healthy for humans to have, so being able to target only the illness-causing bacteria is less destructive to the health of our bodies. Knowing the host range of a phage means that it can be used to target those bacteria species in medicine.
Great job Bec! You should be proud of all that you’ve accomplished this semester
thank you!