Thanks for the question Garrison! The first step I would take would be confirming both homoimmunity and phage release in this actual phage. These have been observed in all other M cluster phages studied but not yet Auspice because those tests haven’t been conducted yet. Homoimmunity is when the phage genome has incorporated into that of the host and other Auspice phages are prevented from lysing the infected host. Phage release is when environmental factors cause the phage to exit lysogeny and kill its host cell. If Auspice shows capable of both, I would then see if the removal or inhibition of any of the proposed genes affect the phage’s ability to create prophage or exit lysogeny. For example if genes 1&2 play the role I proposed we would expect a sample of Auspice with those genes removed to be unable to form functional lysogens.
Sure! There are thousands of new phages that are isolated every year but only about 20% actually have their genome sequenced because sequencing is quite expensive. Of those we classify them into groups called clusters, which are phages that share more than 50% genetic similarity. Usually clusters are signified by a single letter, in this case M. Annotated genomes are genomes that have been reviewed by researchers and have been processed through bioinformatics software to determine where the individual genes in that sequence are and what their functions may be.
To be honest it’s impossible to know whether there even is a large scale application of the identification of this gene, it’s more that we know something is serving this function from experimental data, but we don’t know which gene is responsible. If it’s being done by a gene serving multiple functions, such as the metallophophoesterase gene proposed in this research, it could expand our knowledge about how these genes evolve and enable us to better understand the role they play for the organism. This could in turn allow us to create safer or more effective phage therapies.
PCR didn’t play a direct role in this research because this wasn’t a wet lab so we didn’t do any actual experiments. In future experiments PCR could be used to confirm whether a gene had successfully been removed or not.
What would be the basic structure and research design to further analyze these genes and their functions?
Thanks for the question Garrison! The first step I would take would be confirming both homoimmunity and phage release in this actual phage. These have been observed in all other M cluster phages studied but not yet Auspice because those tests haven’t been conducted yet. Homoimmunity is when the phage genome has incorporated into that of the host and other Auspice phages are prevented from lysing the infected host. Phage release is when environmental factors cause the phage to exit lysogeny and kill its host cell. If Auspice shows capable of both, I would then see if the removal or inhibition of any of the proposed genes affect the phage’s ability to create prophage or exit lysogeny. For example if genes 1&2 play the role I proposed we would expect a sample of Auspice with those genes removed to be unable to form functional lysogens.
Could you explain what annotated M cluster genomes are?
Sure! There are thousands of new phages that are isolated every year but only about 20% actually have their genome sequenced because sequencing is quite expensive. Of those we classify them into groups called clusters, which are phages that share more than 50% genetic similarity. Usually clusters are signified by a single letter, in this case M. Annotated genomes are genomes that have been reviewed by researchers and have been processed through bioinformatics software to determine where the individual genes in that sequence are and what their functions may be.
What larger implications does the possible presence of RDF gene have?
To be honest it’s impossible to know whether there even is a large scale application of the identification of this gene, it’s more that we know something is serving this function from experimental data, but we don’t know which gene is responsible. If it’s being done by a gene serving multiple functions, such as the metallophophoesterase gene proposed in this research, it could expand our knowledge about how these genes evolve and enable us to better understand the role they play for the organism. This could in turn allow us to create safer or more effective phage therapies.
What crucial is PCR to your research?
PCR didn’t play a direct role in this research because this wasn’t a wet lab so we didn’t do any actual experiments. In future experiments PCR could be used to confirm whether a gene had successfully been removed or not.