Start Date
27-4-2023 10:30 AM
Document Type
Poster
Description
This project will generate reliable data about enzyme and gene functions to allow us to improve existing artificial intelligence models. FoldIt is a modeling software used to predict the impact of mutations using its database. Bacteria E. coli are used to produce Beta-glucosidase B (BglB) proteins as they provide a sound model system because they are populations we understand and can control in the lab, and also provide a template for enzyme mutations. During this process, mutagenesis and sequencing was a success, as the mutation that occurred was on N293, and it mutated to N293M on all three samples done. Using the Kunkel Mutagenesis approach, plasmid pET29b+ -BglB will be used in introducing mutations into protein-coding sequences. After the mutation occurs in the plasmid, annealing, and polymerization are done to create a hybrid plasmid. When the hybrid plasmid is introduced into bacterial cells, the cells break down the original DNA strand and build a new double-stranded mutated version of the plasmid. This plasmid will be used to produce a mutant protein. We will have enzyme activity and stability data from labs to learn the impact of the mutation on enzyme function.
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Mutations and Proteins: Using Models to Create Reliable Information
This project will generate reliable data about enzyme and gene functions to allow us to improve existing artificial intelligence models. FoldIt is a modeling software used to predict the impact of mutations using its database. Bacteria E. coli are used to produce Beta-glucosidase B (BglB) proteins as they provide a sound model system because they are populations we understand and can control in the lab, and also provide a template for enzyme mutations. During this process, mutagenesis and sequencing was a success, as the mutation that occurred was on N293, and it mutated to N293M on all three samples done. Using the Kunkel Mutagenesis approach, plasmid pET29b+ -BglB will be used in introducing mutations into protein-coding sequences. After the mutation occurs in the plasmid, annealing, and polymerization are done to create a hybrid plasmid. When the hybrid plasmid is introduced into bacterial cells, the cells break down the original DNA strand and build a new double-stranded mutated version of the plasmid. This plasmid will be used to produce a mutant protein. We will have enzyme activity and stability data from labs to learn the impact of the mutation on enzyme function.
Comments
The faculty mentor for this project was Heather Seitz, Biology.