Expression of the ECP-2 C-Terminus in Latrodectus Hesperus
Poster Number
17
Format
Poster Presentation
Faculty Mentor Name
Craig Vierra
Faculty Mentor Department
Biological Sciences
Abstract/Artist Statement
Spider silk’s high tensile strength and elasticity, as well as its biocompatibility, can potentially be used to revolutionize medicine and technology. Current research is focused on elucidating the silk manufacturing process and determining a means for mass production of the silk. The egg case protein 2 (ECP-2) was isolated from the egg case silk fibers from the black widow spider, Latrodectus hesperus, and its cDNA sequence was determined using reverse genetics. ECP-2 is 826 amino acid residues long and has a fibroin like structure that contains a highly conserved N-terminus as well as a C-terminus rich in GA repeat residues. ECP-2’s location suggests that it acts as a wrapper or casing for the fibers. The ECP-2 cDNA was amplified using PCR and then placed into the prokaryotic expression vector pBAD-TOPO. The amplified segment corresponded to the C-terminus of the full-length ECP-2 protein. After ligation, the vector was transformed into Escherichia coli. Cells carrying the expression vector were induced and ECP-2 levels were analyzed using a western blot. Following the confirmation of ECP-2 expression, we plan to express and purify large amounts of the ECP-2 C-terminus for structural studies.
Location
DeRosa University Center, Ballroom
Start Date
21-4-2011 6:00 PM
End Date
21-4-2011 8:00 PM
Expression of the ECP-2 C-Terminus in Latrodectus Hesperus
DeRosa University Center, Ballroom
Spider silk’s high tensile strength and elasticity, as well as its biocompatibility, can potentially be used to revolutionize medicine and technology. Current research is focused on elucidating the silk manufacturing process and determining a means for mass production of the silk. The egg case protein 2 (ECP-2) was isolated from the egg case silk fibers from the black widow spider, Latrodectus hesperus, and its cDNA sequence was determined using reverse genetics. ECP-2 is 826 amino acid residues long and has a fibroin like structure that contains a highly conserved N-terminus as well as a C-terminus rich in GA repeat residues. ECP-2’s location suggests that it acts as a wrapper or casing for the fibers. The ECP-2 cDNA was amplified using PCR and then placed into the prokaryotic expression vector pBAD-TOPO. The amplified segment corresponded to the C-terminus of the full-length ECP-2 protein. After ligation, the vector was transformed into Escherichia coli. Cells carrying the expression vector were induced and ECP-2 levels were analyzed using a western blot. Following the confirmation of ECP-2 expression, we plan to express and purify large amounts of the ECP-2 C-terminus for structural studies.