The Structural Studies of Artificial Silk Fibers and the PySp2 Protein

Spider silk is a biodegradable, non-toxic biopolymer that is stronger than Kelvar, Nylon, and steel. Spider silk can be used in a variety of fields, including engineering and medicine. In our research, we are attempting to spin synthetic spider silk from a glue silk protein, PySp2. PySp2, which is expressed in golden orb weavers, is spun into attachment discs and helps immobilize dragline threads. PySp2 contains internal block repeats whose sequences can be tested for their unique mechanical properties. Using genetic engineering, we inserted a segment of the PySp2 cDNA into the prokaryotic expression vector pBAD-Thio-TOPO. Restriction digestion analysis and agarose gel electrophoresis was performed to verify the presence and directionality of the PySp2 cDNA in the cloning vector. Following the confirmation of the cDNA insert in the cloning vector, we induced the expression of PySp2 in bacteria and monitored its expression using western blot analysis. Our long term goal is to purify the PySp2 protein and spin artificial silk fibers as well as use the solubilized protein for structural studies