Pea Seedling Amine Oxidase: Production and Purification
Start Date
7-8-2026 11:45 AM
End Date
7-8-2026 12:00 PM
Location
ALT 203
Abstract
The goal of the summer research was to make pea seedling amine oxidase (PSAO) protein using two different methods. One method was to extract and purify this protein from its native source – pea seedlings. Pea seedlings were ground up and the protein was purified using ammonium sulfate and chromatography procedures. Results from protein gels and protein concentrations showed that very little protein was obtained. Since we need more protein for our experiments, we will need to grow more pea seedlings to get more protein. Another method to obtain this protein was to express this protein in a different plant – Nicotiana benthamiana. This expression system would allow us to make more protein more quickly since we would not need to rely on natural expression levels of the protein. N. benthamiana is an Australian cousin of American tobacco, which has been shown to be useful for expressing different proteins. To express a protein in plants, a DNA vector needs to be constructed containing the gene for the protein as well as other genes needed for the protein to be expressed. The gene and the plasmid are prepared by PCR and ligated using recombinational cloning. We have amplified the gene for PSAO and are working on preparing the plasmid to receive the gene. We have also begun growing N. benthamiana for future protein expression.
Pea Seedling Amine Oxidase: Production and Purification
ALT 203
The goal of the summer research was to make pea seedling amine oxidase (PSAO) protein using two different methods. One method was to extract and purify this protein from its native source – pea seedlings. Pea seedlings were ground up and the protein was purified using ammonium sulfate and chromatography procedures. Results from protein gels and protein concentrations showed that very little protein was obtained. Since we need more protein for our experiments, we will need to grow more pea seedlings to get more protein. Another method to obtain this protein was to express this protein in a different plant – Nicotiana benthamiana. This expression system would allow us to make more protein more quickly since we would not need to rely on natural expression levels of the protein. N. benthamiana is an Australian cousin of American tobacco, which has been shown to be useful for expressing different proteins. To express a protein in plants, a DNA vector needs to be constructed containing the gene for the protein as well as other genes needed for the protein to be expressed. The gene and the plasmid are prepared by PCR and ligated using recombinational cloning. We have amplified the gene for PSAO and are working on preparing the plasmid to receive the gene. We have also begun growing N. benthamiana for future protein expression.
