Poster Presentation HUPO 2019 - 18th Human Proteome Organization World Congress

Biological implications of gene modularisation in the virus φX174 (#920)

Bradley W Wright 1 , Paul R Jaschke 1 , Mark P Molloy 2
  1. Macquarie University, North Ryde, NSW, Australia
  2. Kolling Institute, Sydney University, Sydney, NSW, Australia

Modularisation of native genomic coding elements has been a feature in recent synthetic biology designs. The rationales behind these efforts is to produce gene pathways more amenable to manipulation, and to facilitate more predictable and efficient genome engineering outcomes. However, efforts can be hampered by the discovery of overlapping genes, leading to time and resource intensive characterisation and redesign to off-put losses in system efficiency and fidelity as a result of their disruption. To uncover the biological impacts of gene modularisation we characterise a synthetic φX174 virus (termed decompressed) in which all instances of gene overlap has been removed. Our results indicate that despite decompressed possessing the same coding repertoire as the wild-type, there are deficiencies in attachment, progeny production, and capsid stability. Furthermore, preliminary work at the proteome level through the use of targeted mass spectrometry has facilitated for the first time, identification of all known phage proteins. This will enable us to quantitatively measure and compare phage protein production between the wild-type and its genome decompressed variant. We aim to identify the causative reasons for loss in viability as a result of modularisation through the linkage of protein expression, and phenotype measurements to that of in silico genome structure, and translation rate predictions. We will provide preliminary results of the above.