Exploration of Pyriform Spidroin1 (PySp1) QEA Module’s Role in Fiber Formation and Properties

Poster Number

63

Lead Author Major

Biological Sciences

Format

Poster Presentation

Faculty Mentor Name

Craig Vierra

Faculty Mentor Department

Biological Sciences

Abstract/Artist Statement

Spider silk, known for its tensile strength, extensibility and toughness, has a wide array of potential uses that span from medical equipment to military armor. Spider silk’s strength is comparable to high-tensile steel, yet it can withstand considerable strain or extensibility without breaking. There are several types of spider silk, each originating from a different silk- producing gland. These fibers provide different functions for spiders, ranging from web constructions to locomotion. Pyriform spidroin 1 (PySp1), the protein under study, was first identified in the pyriform gland, whose main function is to produce silk that is used to immobilize dragline silk. Our research focused on the significance of PySp1 QEA module and its relationship to the mechanical adhesive properties of pyriform silks. The PySp1 cDNA encoding the QEA module was amplified from a cDNA library prepared from the silk-producing glands of the black widow spider, Lactrodectus hesperus. After insertion of the cDNA into a prokaryotic expression vector, we transformed E. coli and performed a restriction digestion analysis to check for the cDNA insert presence and correct directionality. Following the validation of the presence of a cDNA insert, the QEA PySp1 cDNA was induced and its product monitored through both blue silver staining and western blotting. Western blot analysis showed that QEA PySp1 was expressed in high levels. To further understand its role in pyriform silk formation and function, we plan to purify the protein through nickel affinity chromatography. We hope to spin fibers from the purified protein solution in order to better comprehend its mechanical properties and function.

Location

Grave Covell

Start Date

21-4-2012 10:00 AM

End Date

21-4-2012 12:00 PM

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Apr 21st, 10:00 AM Apr 21st, 12:00 PM

Exploration of Pyriform Spidroin1 (PySp1) QEA Module’s Role in Fiber Formation and Properties

Grave Covell

Spider silk, known for its tensile strength, extensibility and toughness, has a wide array of potential uses that span from medical equipment to military armor. Spider silk’s strength is comparable to high-tensile steel, yet it can withstand considerable strain or extensibility without breaking. There are several types of spider silk, each originating from a different silk- producing gland. These fibers provide different functions for spiders, ranging from web constructions to locomotion. Pyriform spidroin 1 (PySp1), the protein under study, was first identified in the pyriform gland, whose main function is to produce silk that is used to immobilize dragline silk. Our research focused on the significance of PySp1 QEA module and its relationship to the mechanical adhesive properties of pyriform silks. The PySp1 cDNA encoding the QEA module was amplified from a cDNA library prepared from the silk-producing glands of the black widow spider, Lactrodectus hesperus. After insertion of the cDNA into a prokaryotic expression vector, we transformed E. coli and performed a restriction digestion analysis to check for the cDNA insert presence and correct directionality. Following the validation of the presence of a cDNA insert, the QEA PySp1 cDNA was induced and its product monitored through both blue silver staining and western blotting. Western blot analysis showed that QEA PySp1 was expressed in high levels. To further understand its role in pyriform silk formation and function, we plan to purify the protein through nickel affinity chromatography. We hope to spin fibers from the purified protein solution in order to better comprehend its mechanical properties and function.