Date of Award


Degree Type


Degree Name

Master of Science (MS)


Microbiology and Immunology


Coronaviruses (CoVs) can cause a range of symptoms; from a mild common cold to life threatening acute respiratory distress syndrome (ARDS) upon infection. These emerging viruses have the ability to be highly pathogenic and detrimental to the human population. Two prime examples of CoV emergence, SARS-CoV (Severe Acute Respiratory Syndrome) and MERS-CoV, (Middle Eastern Respiratory Syndrome), which exhibits the pandemic potential of emerging CoVs that gain human tropism. Given the increasing potential of an emerging CoV outbreak there is a state of urgency to develop vaccines that will help protect the human population against current and future circulating strains. The Coronavirdae family of viruses are unique because they contain a highly conserved gene arrangement and function within its viral genome. The purpose of this study is to identify a technique that will target attenuating mutations that are present within a functionally conserved region. The way in which I identified attenuating mutations is through temperature-sensitive phenotyping (TS). Using this technique I was able to identify mutants strains of CoVs that contained TS mutations. This TS phenotype was identified by a reduction in titer and a reduction in plaque size when incubated in non-permissive conditions. By using genomic deep sequencing, I was able to identify novel attenuating mutations in the CoV genome. The results gained from this study confirmed the use of TS phenotyping and sequencing as an advantageous tool to identify sites that confer mutations. Identification of these attenuating mutations allows for the application to current strains and confer potential but also may attenuate future emerging strains as well.

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.

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