Exposure of bacteria (with integrated phage DNA-prophage) to stressors like UV light and low nutrients availability for phages cause phages to excise itself from bacteria chromosome and undergo lytic cycle. When such scenario happens, the phage has been induced to undergo lytic cycle.
The phage I extracted was a temperate phage. Temperate phage are phages that can undergo lytic cycle and Lysogenic cycle that is its plaque formation on an L-agar plate (when plaque assay is performed) will be a clear region if phage undergoes lytic cycle and a turbid halo region with a clear zone in the middle when phage undergo lysogenic cycle. The sizes of phage particles were determined using Electron Microscopy. Unfortunately, I did not get the opportunity to do the EM images so the EM imaging Personnel provided the sizes (tail length) for this phage.
The bacteria used in this research was Mycobacterium smegmatis so the different lawns of bacteria that Riggy could be plated on can be Mycobacterium that are closely related to M. smeg. (example M. goodii, M. alvei, and M. abscessus).
The method that can be used to change a temperate phage to make it a better candidate for phage therapy is to repress the transcription of lysogenic genes (Recombination genes- genes that code for integrase and excisionase). By repressing the recombination genes, the phage can undergo lytic cycle and hence become a good candidate for phage therapy.
The integration of viral DNA in a bacterial cell is not a viable option because we want the phage to lyse the bacteria. In phage therapy, phages are used to cure bacterial infection so if a phage DNA gets integrated in a bacteria chromosome, the bacteria infection will not be cured but rather acquire immunity from the Phage since viral DNA has been integrated into bacteria chromosome.
What makes conditions unfavorable for theses phages?
Exposure of bacteria (with integrated phage DNA-prophage) to stressors like UV light and low nutrients availability for phages cause phages to excise itself from bacteria chromosome and undergo lytic cycle. When such scenario happens, the phage has been induced to undergo lytic cycle.
What kind of phage have you extracted that are shown in the EM, Siphoviridae or Myoviridae? How did you determine the sizes of your phage particles.
The phage I extracted was a temperate phage. Temperate phage are phages that can undergo lytic cycle and Lysogenic cycle that is its plaque formation on an L-agar plate (when plaque assay is performed) will be a clear region if phage undergoes lytic cycle and a turbid halo region with a clear zone in the middle when phage undergo lysogenic cycle. The sizes of phage particles were determined using Electron Microscopy. Unfortunately, I did not get the opportunity to do the EM images so the EM imaging Personnel provided the sizes (tail length) for this phage.
What “different lawns of bacteria” do you think Riggy could be plated on?
The bacteria used in this research was Mycobacterium smegmatis so the different lawns of bacteria that Riggy could be plated on can be Mycobacterium that are closely related to M. smeg. (example M. goodii, M. alvei, and M. abscessus).
hi! What are the methods that can be used to change the phages and make them better candidates for phage therapy
The method that can be used to change a temperate phage to make it a better candidate for phage therapy is to repress the transcription of lysogenic genes (Recombination genes- genes that code for integrase and excisionase). By repressing the recombination genes, the phage can undergo lytic cycle and hence become a good candidate for phage therapy.
hi! what methods could have been used that would alter the phages and make them better for phage therapy
Hi
Why is integration of the virus DNA with the bacterial cell not a viable option for phage therapy?
The integration of viral DNA in a bacterial cell is not a viable option because we want the phage to lyse the bacteria. In phage therapy, phages are used to cure bacterial infection so if a phage DNA gets integrated in a bacteria chromosome, the bacteria infection will not be cured but rather acquire immunity from the Phage since viral DNA has been integrated into bacteria chromosome.
Why is integration of Virus DNA with the bacterial cell not a viable option for phage therapy?