Abstract Title
Phage Engineering to Understand Virus Host Range
Additional Funding Sources
The project described was supported by a student grant from the UI Office of Undergraduate Research. The project described was supported by the Idaho EPSCoR Program through the National Science Foundation under Award No. OIA-1736253. The project described was supported by the NIH COBRE Grant P20GM104420 and the R01 Grant GM076040. The project described was supported by the BEACON Grant DBI-0939454. The project described was supported by the Brian and Gayle Hill Undergraduate Research Fellowship administered by the College of Science at the University of Idaho. The project described was supported by the Karen Pohl Award administered by the Department of Biological Sciences at the University of Idaho.
Abstract
I am examining how mutations in the F protein of ΦX174 affect LPS host specificity.
ΦX174 is a bacteriophage used as a model virus due to its small genome and ease of cultivation in laboratory conditions. The Miller-Wichman Lab has used ΦX174 extensively in experimental evolution studies to address statistical questions about the dynamics of adaptive evolution. The lab is now able to engineer thousands of mutations into the ΦX174 genome using a high-throughput method called saturation mutagenesis. For the first part of my project, I am growing a mutant library of the G on its lab host Escherichia coli C to determine the relative fitness of the mutants. To do this, I am using a high-throughput measure of viral fitness the lab has been developing that involves making thousands of mutations to the virus, growing the mutated viruses in a flask together, and sequencing the DNA of the viruses at different time points to see which variants grow best. To validate this type of measurement, I have performed this method with a mutant library of the G protein and am currently comparing my data to traditional fitness data already collected for certain G protein mutants. I have started growing ΦX174 with mutations in the F protein on nine different non-susceptible hosts with a variety of LPS structures. I will perform the fitness assay on isolated mutants from the LPS variant hosts. The lab's goal is the understand the genetic mechanisms of viral attenuation. This kind of study can have a wider impact on preventing virus outbreaks, designing vaccines, or even experimental treatments using bacteriophage in place of antibiotics.
Phage Engineering to Understand Virus Host Range
I am examining how mutations in the F protein of ΦX174 affect LPS host specificity.
ΦX174 is a bacteriophage used as a model virus due to its small genome and ease of cultivation in laboratory conditions. The Miller-Wichman Lab has used ΦX174 extensively in experimental evolution studies to address statistical questions about the dynamics of adaptive evolution. The lab is now able to engineer thousands of mutations into the ΦX174 genome using a high-throughput method called saturation mutagenesis. For the first part of my project, I am growing a mutant library of the G on its lab host Escherichia coli C to determine the relative fitness of the mutants. To do this, I am using a high-throughput measure of viral fitness the lab has been developing that involves making thousands of mutations to the virus, growing the mutated viruses in a flask together, and sequencing the DNA of the viruses at different time points to see which variants grow best. To validate this type of measurement, I have performed this method with a mutant library of the G protein and am currently comparing my data to traditional fitness data already collected for certain G protein mutants. I have started growing ΦX174 with mutations in the F protein on nine different non-susceptible hosts with a variety of LPS structures. I will perform the fitness assay on isolated mutants from the LPS variant hosts. The lab's goal is the understand the genetic mechanisms of viral attenuation. This kind of study can have a wider impact on preventing virus outbreaks, designing vaccines, or even experimental treatments using bacteriophage in place of antibiotics.