Presentation Title

GENETIC SYSTEM CONSTRUCTION OF STIV3: A NEW LYSOGENIC VARIANT OF SULFOLOBUS TURRETED ICOSAHEDRAL VIRUS

Faculty Mentor

Jamie C Snyder

Start Date

23-11-2019 10:45 AM

End Date

23-11-2019 11:30 AM

Location

90

Session

poster 4

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

Despite infecting an entire domain of life, little is known about archaeal viruses and their replication cycle. This is in part due to the lack of genetic systems available to conduct studies into gene and protein function of these biological entities. STIV3 was first isolated as a provirus within its host Sulfolobus acidocaldarius during sequencing of isolates from Yellowstone National Park (YNP) acidic hot springs. It was found to be a viral variant of STIV, as it was observed to be lysogenic and infects a different host with a more stable genome. A genetic system for this virus was constructed by partitioning the genome into five distinct subclones through PCR and ligation into the Escherichia coli cloning vector pSmart. These five subclones were then combined into an infectious clone using the NEBuilder HIFI assembly system that can be replicated in E. coli. Upon electroporation of the STIV3 infectious clone (-pSmart) into its host, free virions were produced that had the ability to infect a different culture of S. acidocaldarius. Work is currently being done for site-directed mutagenesis of genes within each of the subclones to establish its efficacy. This genetic system holds a strong foundation to begin future studies as it will help us to understand lysogeny and gene function in archaeal viruses as well as allow us to use established genetic tools in S. acidocaldarius.

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Nov 23rd, 10:45 AM Nov 23rd, 11:30 AM

GENETIC SYSTEM CONSTRUCTION OF STIV3: A NEW LYSOGENIC VARIANT OF SULFOLOBUS TURRETED ICOSAHEDRAL VIRUS

90

Despite infecting an entire domain of life, little is known about archaeal viruses and their replication cycle. This is in part due to the lack of genetic systems available to conduct studies into gene and protein function of these biological entities. STIV3 was first isolated as a provirus within its host Sulfolobus acidocaldarius during sequencing of isolates from Yellowstone National Park (YNP) acidic hot springs. It was found to be a viral variant of STIV, as it was observed to be lysogenic and infects a different host with a more stable genome. A genetic system for this virus was constructed by partitioning the genome into five distinct subclones through PCR and ligation into the Escherichia coli cloning vector pSmart. These five subclones were then combined into an infectious clone using the NEBuilder HIFI assembly system that can be replicated in E. coli. Upon electroporation of the STIV3 infectious clone (-pSmart) into its host, free virions were produced that had the ability to infect a different culture of S. acidocaldarius. Work is currently being done for site-directed mutagenesis of genes within each of the subclones to establish its efficacy. This genetic system holds a strong foundation to begin future studies as it will help us to understand lysogeny and gene function in archaeal viruses as well as allow us to use established genetic tools in S. acidocaldarius.