Expression of Spider Egg Case Protein-3 for Structural and Mechanical Studies
Poster Number
67
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
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.