Characterization of a new egg case protein from the black widow spider, Latrodectus hesperus.
Poster Number
16
Format
Poster Presentation
Faculty Mentor Name
Craig Vierra
Abstract/Artist Statement
Spider silk is an extremely strong material and material scientists have demonstrated that silk is five times stronger than steel. It has been suggested that a pencil thick strand of silk could stop a Boeing 747 in flight. Because of its high tensile strength and biodegradability, scientists have taken particular interest in the expression, assembly, and spinning processes of silk in order to better serve the environment in the future. By using a eDNA library prepared from the silkproducing glands of the black widow spider, Latrodectus hesperus, primers were designed to amplify a novel gene called egg case silk protein 3 (ECP-3). The expression pattern for ECP-3 is restricted to the tubuliform gland, a gland that is responsible for manufacturing the large diameter fibers of egg case silk. Using PCR, the ECP-3 eDNA was amplified, then gel purified, followed by ligation into a bacterial expression vector. Restriction digestion analysis confirmed the ECP-3 eDNA was in the correct orientation in the cloning vector. Following the verification of the insert in the cloning vector, we induced the expression ofECP-3 in bacteria and verified its expression using western blot analysis. Our long-term goal is to produce the protein in large quantities and purify it from bacteria. This purified protein will then be used to generate anti-ECP-3 antibodies in rabbits, which then can be used as an immunological reagent for studying the molecular properties ofECP-3 in the black widow spider fibers.
Location
Wendell Phillips Center, 1st floor hallways
Start Date
3-5-2008 1:00 PM
End Date
3-5-2008 3:00 PM
Characterization of a new egg case protein from the black widow spider, Latrodectus hesperus.
Wendell Phillips Center, 1st floor hallways
Spider silk is an extremely strong material and material scientists have demonstrated that silk is five times stronger than steel. It has been suggested that a pencil thick strand of silk could stop a Boeing 747 in flight. Because of its high tensile strength and biodegradability, scientists have taken particular interest in the expression, assembly, and spinning processes of silk in order to better serve the environment in the future. By using a eDNA library prepared from the silkproducing glands of the black widow spider, Latrodectus hesperus, primers were designed to amplify a novel gene called egg case silk protein 3 (ECP-3). The expression pattern for ECP-3 is restricted to the tubuliform gland, a gland that is responsible for manufacturing the large diameter fibers of egg case silk. Using PCR, the ECP-3 eDNA was amplified, then gel purified, followed by ligation into a bacterial expression vector. Restriction digestion analysis confirmed the ECP-3 eDNA was in the correct orientation in the cloning vector. Following the verification of the insert in the cloning vector, we induced the expression ofECP-3 in bacteria and verified its expression using western blot analysis. Our long-term goal is to produce the protein in large quantities and purify it from bacteria. This purified protein will then be used to generate anti-ECP-3 antibodies in rabbits, which then can be used as an immunological reagent for studying the molecular properties ofECP-3 in the black widow spider fibers.