Why would genome sequencing allow you to determine which bacterium your phage would be best at infecting? And is there anything else genome sequencing would allow you to determine?
Genome sequencing would help us to categorize our phage into a “cluster” of closely related phages. By observing what bacterium the other phages in that cluster can infect, we can gain insight into what species of bacteria our phage may be able to infect. Genome sequencing can also help us understand how our phage may have evolved or mutated from similar phages, and what genes/proteins/enzymes it uses to function. To see examples of phages that have had their genomes sequenced check out phamerator.org and look at the genome maps. They’re super cool!
Hey! This was a great presentation. I am wondering how you would test this phage in the later steps of antibiotic development? How does it impact human cells and how would it be introduced into the body to treat an infection?
Hey Ava great question! In the later stages of developing this phage into phage therapy, we would use dilutions and spot tests to determine an appropriate dosage to be used. The dosage could then be tested in mice or other animals to make sure it is safe before testing it in humans. One concern on the impact of phage therapy is causing an autoimmune response that could be fatal. The phage therapy could be introduced into the body intravenously or through an inhaler depending on where the pathogen is affecting the patient.
We used many procedures over several weeks to isolate our phage from the soil sample. We first enriched the soil with M. smegmatis, phage buffer, and LB media to allow the phage to replicate and survive. Then we centrifuged and filtered the sample before plating the phage on an agar plate. Once we saw plaques on the plates we scraped off the plaques to obtain the phage. Then we further diluted and filtered the sample to ensure we isolated the phage.
Hi Olya! Great question. Antibiotics have been around for decades and not changed much, and they are also very overprescribed. Bacteria can replicate and mutate very quickly, and when bacteria are exposed to antibiotics, only the bacteria with a mutation that makes them resistant to the antibiotics will survive. Antibiotic resistant bacteria will spawn more antibiotic bacteria as they replicate. This causes the effectiveness of antibiotics to decrease on that population of bacteria. We call this “antibiotic resistance”. Due to this issue, as time goes on our current antibiotics will only become less and less effective, so scientists are looking for an alternative!
Why would genome sequencing allow you to determine which bacterium your phage would be best at infecting? And is there anything else genome sequencing would allow you to determine?
Genome sequencing would help us to categorize our phage into a “cluster” of closely related phages. By observing what bacterium the other phages in that cluster can infect, we can gain insight into what species of bacteria our phage may be able to infect. Genome sequencing can also help us understand how our phage may have evolved or mutated from similar phages, and what genes/proteins/enzymes it uses to function. To see examples of phages that have had their genomes sequenced check out phamerator.org and look at the genome maps. They’re super cool!
Hey! This was a great presentation. I am wondering how you would test this phage in the later steps of antibiotic development? How does it impact human cells and how would it be introduced into the body to treat an infection?
Hey Ava great question! In the later stages of developing this phage into phage therapy, we would use dilutions and spot tests to determine an appropriate dosage to be used. The dosage could then be tested in mice or other animals to make sure it is safe before testing it in humans. One concern on the impact of phage therapy is causing an autoimmune response that could be fatal. The phage therapy could be introduced into the body intravenously or through an inhaler depending on where the pathogen is affecting the patient.
What methods did you use to isolate the phage from the soil?
We used many procedures over several weeks to isolate our phage from the soil sample. We first enriched the soil with M. smegmatis, phage buffer, and LB media to allow the phage to replicate and survive. Then we centrifuged and filtered the sample before plating the phage on an agar plate. Once we saw plaques on the plates we scraped off the plaques to obtain the phage. Then we further diluted and filtered the sample to ensure we isolated the phage.
Why are alternatives to antibiotics necessary?
Hi Olya! Great question. Antibiotics have been around for decades and not changed much, and they are also very overprescribed. Bacteria can replicate and mutate very quickly, and when bacteria are exposed to antibiotics, only the bacteria with a mutation that makes them resistant to the antibiotics will survive. Antibiotic resistant bacteria will spawn more antibiotic bacteria as they replicate. This causes the effectiveness of antibiotics to decrease on that population of bacteria. We call this “antibiotic resistance”. Due to this issue, as time goes on our current antibiotics will only become less and less effective, so scientists are looking for an alternative!