Title

Cracking the Shell

Poster Number

13

Lead Author Major

Biological Sciences

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

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Apr 21st, 6:00 PM Apr 21st, 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.