Title

Bacterial Expression of Fasciclin: A Key Component of Dragline Silk

Poster Number

02B

Lead Author Major

Biology

Lead Author Status

Junior

Second Author Major

Pre-Dentistry

Second Author Status

Sophomore

Third Author Major

Pre-Dentistry

Third Author Status

Sophomore

Fourth Author Major

Biology

Fourth Author Status

Junior

Format

Poster Presentation

Faculty Mentor Name

Craig Vierra

Faculty Mentor Email

cvierra@pacific.edu

Faculty Mentor Department

Biological Sciences

Abstract/Artist Statement

Dragline silk, the primary locomotive silk produced by the black widow spider, can be used for a number of industrial applications, due to its tensile strength, extensibility, and toughness. Its material properties give it considerable versatility, since it has greater tensile strength than high tensile steel and is comparable toughness to Kevlar (body armour). Furthermore, dragline silk is eco-friendly, consisting of organic properties that make it biodegradable. Upon proteomic analysis, dragline silk was found to be composed of 7 different proteins common to the major ampullate gland, the spinning dope, and dragline silk. These proteins include MaSp1, MaSp2, CRISP3, fasciclin, and 3 cysteine rich proteins: CRP1, CRP2, CRP4. This study focused primarily on fasciclin, one of the 7 common proteins. Isolation of the fasciclin cDNA was accomplished by PCR amplification of the gene. The resulting fasciclin cDNA was amplified, extracted via agarose gel electrophoresis, then inserted into a pET-19b SUMO plasmid expression vector. Next, we transformed the expression vector into competent E.coli cells for expression studies. Following purification of fasciclin, mass spectroscopy was used to validate the purification of the protein after affinity purification. Our results demonstrate that we successfully expressed and purified the fasciclin protein from bacterial cells.

Location

DeRosa University Center, Ballroom

Start Date

29-4-2017 1:00 PM

End Date

29-4-2017 3:00 PM

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Apr 29th, 1:00 PM Apr 29th, 3:00 PM

Bacterial Expression of Fasciclin: A Key Component of Dragline Silk

DeRosa University Center, Ballroom

Dragline silk, the primary locomotive silk produced by the black widow spider, can be used for a number of industrial applications, due to its tensile strength, extensibility, and toughness. Its material properties give it considerable versatility, since it has greater tensile strength than high tensile steel and is comparable toughness to Kevlar (body armour). Furthermore, dragline silk is eco-friendly, consisting of organic properties that make it biodegradable. Upon proteomic analysis, dragline silk was found to be composed of 7 different proteins common to the major ampullate gland, the spinning dope, and dragline silk. These proteins include MaSp1, MaSp2, CRISP3, fasciclin, and 3 cysteine rich proteins: CRP1, CRP2, CRP4. This study focused primarily on fasciclin, one of the 7 common proteins. Isolation of the fasciclin cDNA was accomplished by PCR amplification of the gene. The resulting fasciclin cDNA was amplified, extracted via agarose gel electrophoresis, then inserted into a pET-19b SUMO plasmid expression vector. Next, we transformed the expression vector into competent E.coli cells for expression studies. Following purification of fasciclin, mass spectroscopy was used to validate the purification of the protein after affinity purification. Our results demonstrate that we successfully expressed and purified the fasciclin protein from bacterial cells.