Title

Expression of an Internal Block Repeat Domain from Pyriform Spidroin1, a Glue Silk Protein from Black Widow Spiders

Poster Number

65

Lead Author Major

Biological Sciences

Format

Poster Presentation

Faculty Mentor Name

Craig Vierra

Faculty Mentor Department

Biological Sciences

Abstract/Artist Statement

Spider silk is composed of proteins that allow for its high extensibility and tensile strength, making spider silk an ideal substance for stitching wounds, bandaging, and other medical uses. By synthetically producing spider silk proteins in vitro, ample amounts of spider silk can be synthesized for medical purposes more quickly than by natural spider silk production. The Pyriform Spidroin 1 (PySp1) protein of the black widow spider, Latrodectus hesperus, contains an AEQ rich region that participates in fibers that cement dragline silk to solid supports. The mRNA transcripts of PySp1 are expressed and produced in the pyriform gland of the abdomen of the spider. Since PySp1 transcripts are expressed in the pyriform gland and the protein spun into fibers, we hypothesize that PySp1 has an important structural role as it is spun into attachment disc silks to lock dragline fibers to substrates, which is important for prey capture and movement. To produce PySp1 recombinantly, the cDNA was inserted into the prokaryotic expression vector pBAD-Thio-Topo. PySp1 protein induction with arabinose was checked with blue-silver staining and Western blot analysis. The recombinant protein will be purified using a nickel resin and then used to spin synthetic fibers.

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

Expression of an Internal Block Repeat Domain from Pyriform Spidroin1, a Glue Silk Protein from Black Widow Spiders

Grave Covell

Spider silk is composed of proteins that allow for its high extensibility and tensile strength, making spider silk an ideal substance for stitching wounds, bandaging, and other medical uses. By synthetically producing spider silk proteins in vitro, ample amounts of spider silk can be synthesized for medical purposes more quickly than by natural spider silk production. The Pyriform Spidroin 1 (PySp1) protein of the black widow spider, Latrodectus hesperus, contains an AEQ rich region that participates in fibers that cement dragline silk to solid supports. The mRNA transcripts of PySp1 are expressed and produced in the pyriform gland of the abdomen of the spider. Since PySp1 transcripts are expressed in the pyriform gland and the protein spun into fibers, we hypothesize that PySp1 has an important structural role as it is spun into attachment disc silks to lock dragline fibers to substrates, which is important for prey capture and movement. To produce PySp1 recombinantly, the cDNA was inserted into the prokaryotic expression vector pBAD-Thio-Topo. PySp1 protein induction with arabinose was checked with blue-silver staining and Western blot analysis. The recombinant protein will be purified using a nickel resin and then used to spin synthetic fibers.