Presentation Type

Oral Presentation

Start Date

5-8-2024 11:00 AM

End Date

5-8-2024 1:00 PM

Subjects

Mutagenesis, Virology

Advisor

Dr. Kenneth Stedman

Student Level

Undergraduate

Abstract

Archaeal viruses with unique structures such as spindle-shaped virions are found abundantly in extreme environments like geothermal hot springs around the world. Among all spindle-shaped viruses, the model Sulfolobus Spindle-shaped Virus 1 (SSV1) is best studied. Creating the lemon-shaped or spindle-shaped virion structure are two proteins, VP1 as the major capsid protein, and VP3 as the minor capsid protein. The primary structure of VP1 consists of a proteolytic cleavage site at position 66 that is believed to be necessary for virus evolution. Recent studies showed that genetic mutation of the amino acid, glutamate (E) at position 66 in VP1 which is highly conserved throughout all Spindle-shaped viruses, produces significant changes to the lemon shaped structure of SSV1. Here, construction of VP1 mutants was made by introducing Hexahistidine protein affinity tags in order to observe the levels of expression in VP1 of these E66 mutants that produced different capsid morphologies. SSV1 mutants were generated utilizing an exponential mutagenesis polymerase chain reaction protocol via RepliQa to compose the tagged SSV1 VP1 mutants. Following PCR, mutants were treated with KLD treatment, and then transformed.

Creative Commons License or Rights Statement

Creative Commons Attribution 4.0 License
This work is licensed under a Creative Commons Attribution 4.0 License.

Persistent Identifier

https://archives.pdx.edu/ds/psu/41829

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May 8th, 11:00 AM May 8th, 1:00 PM

Construction and Mutagenesis of SSV1 Mutants in Extreme Viruses

Archaeal viruses with unique structures such as spindle-shaped virions are found abundantly in extreme environments like geothermal hot springs around the world. Among all spindle-shaped viruses, the model Sulfolobus Spindle-shaped Virus 1 (SSV1) is best studied. Creating the lemon-shaped or spindle-shaped virion structure are two proteins, VP1 as the major capsid protein, and VP3 as the minor capsid protein. The primary structure of VP1 consists of a proteolytic cleavage site at position 66 that is believed to be necessary for virus evolution. Recent studies showed that genetic mutation of the amino acid, glutamate (E) at position 66 in VP1 which is highly conserved throughout all Spindle-shaped viruses, produces significant changes to the lemon shaped structure of SSV1. Here, construction of VP1 mutants was made by introducing Hexahistidine protein affinity tags in order to observe the levels of expression in VP1 of these E66 mutants that produced different capsid morphologies. SSV1 mutants were generated utilizing an exponential mutagenesis polymerase chain reaction protocol via RepliQa to compose the tagged SSV1 VP1 mutants. Following PCR, mutants were treated with KLD treatment, and then transformed.