Title

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

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May 5th, 1:00 PM May 5th, 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!