Start Date
27-4-2023 12:00 PM
Document Type
Poster
Description
The research aims to generate reliable data about genes and enzyme function to improve AI models. Modeling software (fold it) is used to predict the impact of mutation in Bgl B enzyme by intentionally modifying the structure of the protein by changing one amino acid residue. The mutation made is methionine to Leucine (M221L). Bacteria E.coli is used to produce Bgl B mutant protein. Bacterial cells have a plasmid used as a vector and trick their cells into making proteins from other organisms. Their cells can read genes from different organisms producing proteins coded by these genes since genetic code and DNA structures are identical across the living world. I introduced the Kunkel product plasmid into a strained E.coli bacteria and added antibiotic-resistant kanamycin to separate the bacterial plasmid from my plasmid. The goal is to use E.coli bacteria to replicate the Kunkel product plasmid. After growing bacterial cells in liquid culture, I extracted the plasmid DNA for downstream sequencing to verify that the Kunkel mutagenesis process succeeded, transforming the strained E.coli for protein production. For successful sequencing, I did a DNA concentration Assay (A260) to determine the concentration of the DNA. In the sequencing results, out of three cultures, two had mutations.
Image
M221L Mutagenesis
The research aims to generate reliable data about genes and enzyme function to improve AI models. Modeling software (fold it) is used to predict the impact of mutation in Bgl B enzyme by intentionally modifying the structure of the protein by changing one amino acid residue. The mutation made is methionine to Leucine (M221L). Bacteria E.coli is used to produce Bgl B mutant protein. Bacterial cells have a plasmid used as a vector and trick their cells into making proteins from other organisms. Their cells can read genes from different organisms producing proteins coded by these genes since genetic code and DNA structures are identical across the living world. I introduced the Kunkel product plasmid into a strained E.coli bacteria and added antibiotic-resistant kanamycin to separate the bacterial plasmid from my plasmid. The goal is to use E.coli bacteria to replicate the Kunkel product plasmid. After growing bacterial cells in liquid culture, I extracted the plasmid DNA for downstream sequencing to verify that the Kunkel mutagenesis process succeeded, transforming the strained E.coli for protein production. For successful sequencing, I did a DNA concentration Assay (A260) to determine the concentration of the DNA. In the sequencing results, out of three cultures, two had mutations.
Comments
The faculty mentor for this project was Heather Seitz, Biology.