Yes! We streaked the sample in a way so that the plaques would be pretty evenly spread on the plate, but the placement of the plaques on the plate doesn’t really matter, just the size and if the plaque is clear/cloudy matters
How common are cluster A phages? Is this the most common cluster? Do you have any hypothesis about which sub-cluster your phage may be in since it likely isn’t in A3 or A10?
I don’t have any numbers or statistics on how common A cluster phages are, but there are a lot of A subclusters, so I’m guessing there are a lot of A cluster phages. My next subcluster hypothesis is A1, because phages in this subcluster have similar restriction digest cut patterns, phage morphologies, and plaque morphologies as our phage.
You could engineer a phage to be lytic by repressing the expression of the repressor protein in the temperate phage. Repressor proteins bind to an operator on the phage genome which inhibits the expression of lytic genes. By inhibiting the expression of the repressor protein, the expression of the lytic genes won’t be repressed and the temperate phage would immediately enter the lytic lifecycle.
what does the subclusters A3 and A10 mean? i dont think i understand what a subcluster means in this sense.
Subclusters are just a way to organize/categorize phages. Phages with similar genomes will be placed in the same cluster/subcluster
Do plaque streaks affect where the spots of phage are on the dish?
Yes! We streaked the sample in a way so that the plaques would be pretty evenly spread on the plate, but the placement of the plaques on the plate doesn’t really matter, just the size and if the plaque is clear/cloudy matters
How common are cluster A phages? Is this the most common cluster? Do you have any hypothesis about which sub-cluster your phage may be in since it likely isn’t in A3 or A10?
I don’t have any numbers or statistics on how common A cluster phages are, but there are a lot of A subclusters, so I’m guessing there are a lot of A cluster phages. My next subcluster hypothesis is A1, because phages in this subcluster have similar restriction digest cut patterns, phage morphologies, and plaque morphologies as our phage.
How would you engineer a phage to be lytic if it’s temperate?
You could engineer a phage to be lytic by repressing the expression of the repressor protein in the temperate phage. Repressor proteins bind to an operator on the phage genome which inhibits the expression of lytic genes. By inhibiting the expression of the repressor protein, the expression of the lytic genes won’t be repressed and the temperate phage would immediately enter the lytic lifecycle.