9 thoughts on “P61 – Dodson

  1. What does the B1 group mean for the phage, does it have an effect on the possibility of the phage to have antibiotic resistant strains, or cause it to be part of the lytic cycle rather than the lysogenic cycle?

  2. What does the phage belonging to the B1 group say about the phage that you studied, does it increase its likelihood of being used to treat antibiotic resistance, or does it just classify the type if phage?

    1. B1 subtype is just a way to classify phages that have very similar but not entirely identical genomes and their DNA is all cut similarly by the same restriction enzymes. If we were to fully sequence our phage genome, comparing to other B1 phages that are lytic may help us determine what genes are specifically making our phage temperate and if we can disable them from being expressed! B1 phages are considered the most common type of phage we’ve databased so far, but it doesn’t necessarily increase its likelihood of being used against antibiotic resistant bacteria since ours has temperate genes.

  3. Since your phage is a temperate phage, how you would you go about getting the phage to lytic as you talked about wanting to do for future directions?

    1. One option we could do is to find a way to excise repressor and integrase genes from our phage to prevent it from incorporating into the bacterial genome and staying dormant. This could force it to circularize in the bacterial cell and begin replicating immediately

  4. Was the restriction digest done on the phage samples in their entirety or just on their DNA? If it was just on the DNA, how did you manage to isolate it from the rest of the sample?

    1. Great question! The restriction digest was only done on isolated phage DNA. We used two enzymes, DNAse I and RNAse A, to hydrolize any bacterial DNA but they won’t destroy our phage DNA since it’s stored and protected inside the capsid head

  5. Your conclusion mentions that there’s potential for selective targeting of bacteria, what kind of future methods/tests if any could further develop this idea?

    1. Since phages are highly specific in which bacteria they can infect, one way we could test what bacteria our phage can infect is to perform many different plaque assays with multiple different related bacterial strains to M. smeg and look for plaque formations (indicating infection). Fully sequencing our phage genome may also indicate which genes it’s using to detect and bind to bacterial cell surfaces, and from there we may be able to tell what types of bacteria it selects for.

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