Growth and characterization of sulfolobus turreted icosahedral virus

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Montana State University - Bozeman, College of Engineering


Sulfolobus turreted icosahedral virus (STIV) and its infection cycle in Sulfolobus solfataricus P2 (S. solfataricus P2) was studied. A method to quantify the virus quickly was developed and optimized. The host-virus relationship between STIV and S. solfataricus P2 was studied and the transcripts of the virus were identified. Cultures of S. solfataricus P2 were grown and infected with STIV. Cultures were infected with different amounts of the virus, at various temperatures and pHs, and virus production was monitored. ELISA protocol was developed to quantify the virus. Southern blots were performed to identify if the virus integrates into its host genome and Northern blots were carried out to identify viral transcripts in culture. A condition was found where STIV could be produced consistently and monitored with Enzyme-Linked ImmunoSorbent Assay (ELISA). Viral production is at a maximum when cells were grown at pH 2.5 and in a temperature range of 75-85°C. The magnitude of virus production does not depend on amount of virus used to infect cultures, when the Multiplicity of Infection (MOI) is above 0.45. Southern blotting did show ideal banding pattern for a non-integrated virus, thus no evidence for integration of STIV into its host genome was found. A total of 9 viral transcripts were identified by Northern blotting of STIV infected S. solfataricus P2 cultures.




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