Cracking the Shell
Poster Number
13
Format
Poster Presentation
Faculty Mentor Name
Craig Vierra
Faculty Mentor Department
Biological Sciences
Abstract/Artist Statement
The black widow spider, Latrodectus hesperus, produces seven different silk proteins that can be spun into various kinds of silks. One of these fiber types, called tubuliform silk, has been shown to be composed of at least three different proteins TuSp1, ECP-1 and ECP-2. Tubuliform silks are found in egg sacs and serve to protect spider embryos during development. Analyses of mRNA levels have shown that ECP-2 is expressed at higher levels relative to ECP-1. The ECPs have been hypothesized to constitute the outer layer of the tubuliform silk fibers. To elucidate the structural role of ECP-2, we have attempted to express part of the protein in bacteria. In order to accomplish this task, we amplified a segment of the ECP-2 cDNA coding its N-terminus. The ECP-2 cDNA was amplified using PCR, ligated into a prokaryotic expression vector and transformed into E. coli. Following transformation, the recombinant protein was induced and its expression level was analyzed by western blot analysis. Here we show that the N-terminus of ECP-2 can be expressed in high levels in bacteria, which should make purification of large amounts of ECP-2 for structural analyses feasible.
Location
DeRosa University Center, Ballroom
Start Date
21-4-2011 6:00 PM
End Date
21-4-2011 8:00 PM
Cracking the Shell
DeRosa University Center, Ballroom
The black widow spider, Latrodectus hesperus, produces seven different silk proteins that can be spun into various kinds of silks. One of these fiber types, called tubuliform silk, has been shown to be composed of at least three different proteins TuSp1, ECP-1 and ECP-2. Tubuliform silks are found in egg sacs and serve to protect spider embryos during development. Analyses of mRNA levels have shown that ECP-2 is expressed at higher levels relative to ECP-1. The ECPs have been hypothesized to constitute the outer layer of the tubuliform silk fibers. To elucidate the structural role of ECP-2, we have attempted to express part of the protein in bacteria. In order to accomplish this task, we amplified a segment of the ECP-2 cDNA coding its N-terminus. The ECP-2 cDNA was amplified using PCR, ligated into a prokaryotic expression vector and transformed into E. coli. Following transformation, the recombinant protein was induced and its expression level was analyzed by western blot analysis. Here we show that the N-terminus of ECP-2 can be expressed in high levels in bacteria, which should make purification of large amounts of ECP-2 for structural analyses feasible.