Expression of human SLP-1 protein in the yeast Pichia pastoris
Poster Number
18
Format
Poster Presentation
Abstract/Artist Statement
Pichia pastoris is a methylotrophic yeast that is widely used for its ability to express large quantities of heterologous proteins. P. pastoris has an alcohol oxidase, AOX1, an inducible promoter that allows it to express proteins in the presence of methanol, but not in the presence of glucose. Currently, we are working with P. pastoris to express the functional human SLP-1 protein. This protein is normally found in saliva. In addition to that this protein is believed to have anti- HIV properties.We created an expression vector with the SLP-1 gene and inserted into P. pastoris strain to express high levels of SLP-1 protein. The yeast was grown under conditions that would allow it to optimally produce large amounts of SLP-1.We attempted to purify the SLP-1 protein from Pichia pastoris extracellular medium. Under small scale conditions, the purification was successful. We then tried purifying a large quantity of the protein using two different methods. Our first method, which involved using a Nickel binding column, was a little more difficult to use. We will soon see whether our second method, using a Cobalt binding column, is a winner!
Location
Pacific Geosciences Center
Start Date
5-5-2007 1:00 PM
End Date
5-5-2007 3:00 PM
Expression of human SLP-1 protein in the yeast Pichia pastoris
Pacific Geosciences Center
Pichia pastoris is a methylotrophic yeast that is widely used for its ability to express large quantities of heterologous proteins. P. pastoris has an alcohol oxidase, AOX1, an inducible promoter that allows it to express proteins in the presence of methanol, but not in the presence of glucose. Currently, we are working with P. pastoris to express the functional human SLP-1 protein. This protein is normally found in saliva. In addition to that this protein is believed to have anti- HIV properties.We created an expression vector with the SLP-1 gene and inserted into P. pastoris strain to express high levels of SLP-1 protein. The yeast was grown under conditions that would allow it to optimally produce large amounts of SLP-1.We attempted to purify the SLP-1 protein from Pichia pastoris extracellular medium. Under small scale conditions, the purification was successful. We then tried purifying a large quantity of the protein using two different methods. Our first method, which involved using a Nickel binding column, was a little more difficult to use. We will soon see whether our second method, using a Cobalt binding column, is a winner!