Hello! You mentioned that phage can be classified based on their tails. What exactly is the advantage or disadvantage of a phage that has a flexible tail versus contractile tail? Or a shorter tail versus a longer tail?
A phage’s tail can dictate a couple of different things about the phage itself. A phage’s tail dictates it’s infection efficiency and host range. A flexible tail has a broader host range which means it can infect wider range of bacterial hosts. There is also a greater reach that allows the phage to explore a larger surface area on the bacterial cell (this can enhance the chances of successful attachment and infection). A contractile tail allows rapid and forceful injection into the bacterial cell. This enhances the efficiency of injection. The rapid injection also allows for a faster infection cycle (increasing the phage population faster). A longer tail may be able to reach the bacterial receptors that are farther away from the cell surface, which can increase the changes of successful attachment. They also can provide more stability during the attachment process. However, phages with longer tails may have a narrower host range due to the reliance of specific bacterial surfaces for attachment. Longer tails may also be harder to move, this reduces the ability to locate and infect target bacteria.
Hello! I really enjoyed your presentation. Can you talk a bit about why it’s necessary to run a gel electrophoresis? I understand that doing this can provide information about the quantity and quality of the phage DNA, but I don’t understand if that information is a commentary on the specific sample collected, or if it’s generalizable to the phage being studied (and this is probably just due to my own lack of familiarity with the method). Thank you!
Hello! The gel electrophoresis can provide different information based on what exactly you are looking for as the researcher. In a Quality Control Gel, the gel itself provides information about the quantity and quality of your specific sample collected. After enrichment, spot test, purification, calculating the HTL, and DNA isolation of your specific phage you would then run the quality control gel to determine if the phage DNA is in enough abundance and enough quality to continue on with restriction digest and PCR. If your DNA is not of high quality and quantity you would want to redo DNA isolation, until it is. Typically, everyone running a quality control gel would want the gel to show that the DNA is high in quality and quantity, but with a restriction digest or PCR you would get different results based on your specific sample.
Your mentioned briefly how tagging with florescence could help you to understand the location of the bacterial infection. Any way you could explain more clearly what that means?
Yes, because temperate phage can remain dormant for a period of time, they typically last longer in the body. This makes them a great candidate for locating specific areas in the body where infections may be located. Typically the phage genome is genetically modified to incorporate a suitable fluorescent tag that will emit light of a specific wavelength. The phage will then be introduced into the host bacteria. The infections then will trigger the replication of the phage within the host. As the phage replicated, the gene encoding the fluorescent tag is expressed. This produces a light that can be viewed on a fluorescence microscopy. Which allows scientists to track the movement and distribution of the phage within a bacterial population.
You mentioned Phages can undergo either lytic cycles or Lysogenic cycles, What are the benefits and drawbacks/uses as a phage therapy and how do these support your conclusion that your phage is lytic and a candidate for further research as a potential phage therapy? Thank you! wonderful presentation.
In phage therapy, the scientists would remove the repressor or integrase (or both) protein if the phage is temperate. This would allow the phage to not go through the lysogenic lifestyle, but instead the lytic lifecycle. The lytic lifecycle allows immediate replication and killing of the bacteria instead of the phage being dormant for a period of time. In order for our phage to be a candidate for phage therapy, the repressor protein would have to be removed.
In your presentation you mentioned all the information would be helpful for scientists to “figure all the stuff out,” what are those “stuff”? Also what characteristics would the scientists focus on more for research?
The information we concluded from our experiment allows scientists to further investigate the properties of a phage’s lifecycle, the use of temperate phages in treating bacterial infections, evolutionary relationships between different species of phage, the detection of certain bacteria on different substances or inside different substances, further research investigation in phage, and agricultural benefits. I believe in our specific research scientists should focus on the removal of the proteins (integrase and repressor) in order to transform our temperate phage that undergoes the lysogenic lifecycle into going straight into the lytic lifecycle. Lytic phages are typically used in phage therapy.
Hey Alexa, great presentation. I was just wondering why your ladder (the right most column of your gel) has no bands that align with or overlap with you sample bands?
The DNA ladder was just meant to serve as a reference tool for our quality control gel. We used it in order to estimate the size of the DNA fragments and to ensure the accuracy of the gel that we ran.
Hello! You mentioned that phage can be classified based on their tails. What exactly is the advantage or disadvantage of a phage that has a flexible tail versus contractile tail? Or a shorter tail versus a longer tail?
A phage’s tail can dictate a couple of different things about the phage itself. A phage’s tail dictates it’s infection efficiency and host range. A flexible tail has a broader host range which means it can infect wider range of bacterial hosts. There is also a greater reach that allows the phage to explore a larger surface area on the bacterial cell (this can enhance the chances of successful attachment and infection). A contractile tail allows rapid and forceful injection into the bacterial cell. This enhances the efficiency of injection. The rapid injection also allows for a faster infection cycle (increasing the phage population faster). A longer tail may be able to reach the bacterial receptors that are farther away from the cell surface, which can increase the changes of successful attachment. They also can provide more stability during the attachment process. However, phages with longer tails may have a narrower host range due to the reliance of specific bacterial surfaces for attachment. Longer tails may also be harder to move, this reduces the ability to locate and infect target bacteria.
Hello! I really enjoyed your presentation. Can you talk a bit about why it’s necessary to run a gel electrophoresis? I understand that doing this can provide information about the quantity and quality of the phage DNA, but I don’t understand if that information is a commentary on the specific sample collected, or if it’s generalizable to the phage being studied (and this is probably just due to my own lack of familiarity with the method). Thank you!
Hello! The gel electrophoresis can provide different information based on what exactly you are looking for as the researcher. In a Quality Control Gel, the gel itself provides information about the quantity and quality of your specific sample collected. After enrichment, spot test, purification, calculating the HTL, and DNA isolation of your specific phage you would then run the quality control gel to determine if the phage DNA is in enough abundance and enough quality to continue on with restriction digest and PCR. If your DNA is not of high quality and quantity you would want to redo DNA isolation, until it is. Typically, everyone running a quality control gel would want the gel to show that the DNA is high in quality and quantity, but with a restriction digest or PCR you would get different results based on your specific sample.
Your mentioned briefly how tagging with florescence could help you to understand the location of the bacterial infection. Any way you could explain more clearly what that means?
Yes, because temperate phage can remain dormant for a period of time, they typically last longer in the body. This makes them a great candidate for locating specific areas in the body where infections may be located. Typically the phage genome is genetically modified to incorporate a suitable fluorescent tag that will emit light of a specific wavelength. The phage will then be introduced into the host bacteria. The infections then will trigger the replication of the phage within the host. As the phage replicated, the gene encoding the fluorescent tag is expressed. This produces a light that can be viewed on a fluorescence microscopy. Which allows scientists to track the movement and distribution of the phage within a bacterial population.
You mentioned Phages can undergo either lytic cycles or Lysogenic cycles, What are the benefits and drawbacks/uses as a phage therapy and how do these support your conclusion that your phage is lytic and a candidate for further research as a potential phage therapy? Thank you! wonderful presentation.
In phage therapy, the scientists would remove the repressor or integrase (or both) protein if the phage is temperate. This would allow the phage to not go through the lysogenic lifestyle, but instead the lytic lifecycle. The lytic lifecycle allows immediate replication and killing of the bacteria instead of the phage being dormant for a period of time. In order for our phage to be a candidate for phage therapy, the repressor protein would have to be removed.
In your presentation you mentioned all the information would be helpful for scientists to “figure all the stuff out,” what are those “stuff”? Also what characteristics would the scientists focus on more for research?
The information we concluded from our experiment allows scientists to further investigate the properties of a phage’s lifecycle, the use of temperate phages in treating bacterial infections, evolutionary relationships between different species of phage, the detection of certain bacteria on different substances or inside different substances, further research investigation in phage, and agricultural benefits. I believe in our specific research scientists should focus on the removal of the proteins (integrase and repressor) in order to transform our temperate phage that undergoes the lysogenic lifecycle into going straight into the lytic lifecycle. Lytic phages are typically used in phage therapy.
Hey Alexa, great presentation. I was just wondering why your ladder (the right most column of your gel) has no bands that align with or overlap with you sample bands?
Hey Ben!
The DNA ladder was just meant to serve as a reference tool for our quality control gel. We used it in order to estimate the size of the DNA fragments and to ensure the accuracy of the gel that we ran.