Title

Expression of Spider Egg Case Protein-3 for Structural and Mechanical Studies

Poster Number

67

Lead Author Major

Biological Sciences

Format

Poster Presentation

Faculty Mentor Name

Craig Vierra

Faculty Mentor Department

Biological Sciences

Abstract/Artist Statement

Spiders produce a diverse number of silk proteins that are well-known for their extraordinary mechanical and biological properties after they are spun into fibers. Dragline silk has been the most prominent focus of research because of its exceptional high tensile strength and extensibility. In our research we have focused on the characterization of a third constituent of tubuliform silks (egg case silk). This constituent, which is dubbed Egg Case Protein 3 (ECP-3), is produced exclusively in the tubuliform gland of the black widow spider, Lactrodectus hesperus. The ECP-3 cDNA was obtained from a cDNA library prepared from the silk-producing glands of the spider. This cDNA was placed into the prokaryotic expression vector pBAD-TOPO and then moved into pET-19b-Sumo to facilitate purification. Using western blot analysis, we have verified that the ECP-3 protein is efficiently expressed in bacteria. To test the structural properties of ECP-3, we plan to perform circular dichroism to study the secondary structure of ECP-3. Our long-term goal is to spin artificial silk fibers for mechanical studies to relate the structure of ECP-3 to the material properties of tubuliform silks.

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 Spider Egg Case Protein-3 for Structural and Mechanical Studies

Grave Covell

Spiders produce a diverse number of silk proteins that are well-known for their extraordinary mechanical and biological properties after they are spun into fibers. Dragline silk has been the most prominent focus of research because of its exceptional high tensile strength and extensibility. In our research we have focused on the characterization of a third constituent of tubuliform silks (egg case silk). This constituent, which is dubbed Egg Case Protein 3 (ECP-3), is produced exclusively in the tubuliform gland of the black widow spider, Lactrodectus hesperus. The ECP-3 cDNA was obtained from a cDNA library prepared from the silk-producing glands of the spider. This cDNA was placed into the prokaryotic expression vector pBAD-TOPO and then moved into pET-19b-Sumo to facilitate purification. Using western blot analysis, we have verified that the ECP-3 protein is efficiently expressed in bacteria. To test the structural properties of ECP-3, we plan to perform circular dichroism to study the secondary structure of ECP-3. Our long-term goal is to spin artificial silk fibers for mechanical studies to relate the structure of ECP-3 to the material properties of tubuliform silks.