Protein Extraction/Purification and Cloning Techniques in Dairy Cow Vaccine
Abstract
The dairy industry is faced with the economic challenge of bacterial udder infections. To address this issue we are working on a vaccine that uses bacterial enterotoxins to enable the nasal delivery of purified bacterial antigens. Recombinant vaccinations use genetically engineered DNA so cells will directly produce a desired antigen, resulting in a protective immunological response. The entire cloning process allows a desired gene to be isolated and inserted into a parental vector to form a plasmid that can be transformed into E. coli. To analyze samples collected from the vaccinated cows to check for an antigen-specific response, purified proteins must be made. The two proteins that are extracted and purified for this study are Iron Regulated Surface Determinant A (IsdA) and Clumping Factor A (ClfA) from Staphyloccus aureus. These antigens are used for enzyme-linked immunosorbent assays to measure the concentration of an antibody solution. Purified IsdA and ClfA antigens coat ELISA plates for antibody analysis. Cloning and protein extraction and purification are important techniques used to create a vaccine that could end mastitis in dairy cows.
Protein Extraction/Purification and Cloning Techniques in Dairy Cow Vaccine
The dairy industry is faced with the economic challenge of bacterial udder infections. To address this issue we are working on a vaccine that uses bacterial enterotoxins to enable the nasal delivery of purified bacterial antigens. Recombinant vaccinations use genetically engineered DNA so cells will directly produce a desired antigen, resulting in a protective immunological response. The entire cloning process allows a desired gene to be isolated and inserted into a parental vector to form a plasmid that can be transformed into E. coli. To analyze samples collected from the vaccinated cows to check for an antigen-specific response, purified proteins must be made. The two proteins that are extracted and purified for this study are Iron Regulated Surface Determinant A (IsdA) and Clumping Factor A (ClfA) from Staphyloccus aureus. These antigens are used for enzyme-linked immunosorbent assays to measure the concentration of an antibody solution. Purified IsdA and ClfA antigens coat ELISA plates for antibody analysis. Cloning and protein extraction and purification are important techniques used to create a vaccine that could end mastitis in dairy cows.