Title

Expression of Internal Block Repeats within the PySp2 Protein

Poster Number

66

Lead Author Major

Biological Sciences

Format

Poster Presentation

Faculty Mentor Name

Craig Vierra

Faculty Mentor Department

Biological Sciences

Abstract/Artist Statement

Spider silk is known for its high performance mechanical properties. Spider silk is tougher than high-tensile steel, largely due to the silk’s increased extensibility. The key feature that allows spider silk to possess phenomenal mechanical properties is due to internal block repeats within its protein architecture; these repeats are rich in alanine and glycine. In addition to the internal block repeats, spider silk fibroins all share a non-repetitive N-terminus and C-terminus, along with molecular masses that exceed 250 kDa. Nephila clavipes, commonly referred to as the golden orb weaving spider, has 6-7 silk glands. One of these glands, the pyriform gland, expresses glue silk genes such as PySp2. PySp2 is found in attachment discs and facilitates N. clavipes in locomotion by helping secure dragline silk to solid supports. Pyriform silks are essential for a spider’s survival and this material is spun into a liquid that dries quickly. In an effort to analyze the relationship between the mechanical properties and protein sequence of PySp2, part of the PySp2 cDNA was expressed in bacteria using the prokaryotic expression vector pBAD/Thio-TOPO. The recombinant protein was purified, and then visualized using Coomassie blue staining as well as western blot analysis. The long-term goal of our research is to ultimately spin the purified protein into fibers for mechanical testing.

Location

Grave Covell

Start Date

21-4-2012 10:00 AM

End Date

21-4-2012 12:00 PM

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Apr 21st, 10:00 AM Apr 21st, 12:00 PM

Expression of Internal Block Repeats within the PySp2 Protein

Grave Covell

Spider silk is known for its high performance mechanical properties. Spider silk is tougher than high-tensile steel, largely due to the silk’s increased extensibility. The key feature that allows spider silk to possess phenomenal mechanical properties is due to internal block repeats within its protein architecture; these repeats are rich in alanine and glycine. In addition to the internal block repeats, spider silk fibroins all share a non-repetitive N-terminus and C-terminus, along with molecular masses that exceed 250 kDa. Nephila clavipes, commonly referred to as the golden orb weaving spider, has 6-7 silk glands. One of these glands, the pyriform gland, expresses glue silk genes such as PySp2. PySp2 is found in attachment discs and facilitates N. clavipes in locomotion by helping secure dragline silk to solid supports. Pyriform silks are essential for a spider’s survival and this material is spun into a liquid that dries quickly. In an effort to analyze the relationship between the mechanical properties and protein sequence of PySp2, part of the PySp2 cDNA was expressed in bacteria using the prokaryotic expression vector pBAD/Thio-TOPO. The recombinant protein was purified, and then visualized using Coomassie blue staining as well as western blot analysis. The long-term goal of our research is to ultimately spin the purified protein into fibers for mechanical testing.