Title

Taking the Sticky out of Spider Silk

Poster Number

26

Format

Poster Presentation

Abstract/Artist Statement

Black widow spider silk would be ideal for industrial purposes due to its high tensile strength and elasticity. Its fibers can potentially be used to manufacture different products such as bulletproof vests, sutures, and nanotechnology applications. Unfortunately, native silk proteins are difficult to acquire in substantial amounts due to the dangerous nature of black widow spiders. We are expressing the spider silk protein Pyriform Spidroin 1 (PySp1) in Pichia pastoris, a methylotropic yeast used for heterologous protein expression, in hopes of enabling its synthetic manufacture.The C-terminal region of the spider silk protein was successfully expressed and secreted in large amounts by P. pastoris. The gene for the 40-kD protein was fused to a Myc-tag for easy detection and a His-tag for purification. Isolation of His-tagged protein was attempted under denaturing conditions with urea. Western analysis of the intracellular extracts indicated that the protein aggregated prior to secretion from the cells. Thus, due to this aggregation and the protein's large size, the purification was unsuccessful. We are currently attempting to selectively precipitate the PySp1 proteins with ammonium sulfate and heat. By developing a method to purify PySp1, we will enable scientists to manipulate the spider silk protein for synthetic mass production.

Location

DeRosa University Center, Ballroom B

Start Date

1-5-2010 1:00 PM

End Date

1-5-2010 3:00 PM

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May 1st, 1:00 PM May 1st, 3:00 PM

Taking the Sticky out of Spider Silk

DeRosa University Center, Ballroom B

Black widow spider silk would be ideal for industrial purposes due to its high tensile strength and elasticity. Its fibers can potentially be used to manufacture different products such as bulletproof vests, sutures, and nanotechnology applications. Unfortunately, native silk proteins are difficult to acquire in substantial amounts due to the dangerous nature of black widow spiders. We are expressing the spider silk protein Pyriform Spidroin 1 (PySp1) in Pichia pastoris, a methylotropic yeast used for heterologous protein expression, in hopes of enabling its synthetic manufacture.The C-terminal region of the spider silk protein was successfully expressed and secreted in large amounts by P. pastoris. The gene for the 40-kD protein was fused to a Myc-tag for easy detection and a His-tag for purification. Isolation of His-tagged protein was attempted under denaturing conditions with urea. Western analysis of the intracellular extracts indicated that the protein aggregated prior to secretion from the cells. Thus, due to this aggregation and the protein's large size, the purification was unsuccessful. We are currently attempting to selectively precipitate the PySp1 proteins with ammonium sulfate and heat. By developing a method to purify PySp1, we will enable scientists to manipulate the spider silk protein for synthetic mass production.