Expression of the Latrodectus hesperus Glue Silk Protein, Pyriform Spidroin 1, in Bacteria

The biological mechanisms that spiders use to spin silk fibers remains a mystery to scientists, making it difficult from labs to biomimic this process. Spiders spin multiple silk types that have a diverse range of biological functions. The highly studied dragline silk has been shown to have tremendous properties that rival both steel and Kevlar in elasticity, toughness, and tensile strength. In this lab, the pyriform spidroin protein 1 (PySp1) from the black widow spider, Latrodectus Hesperus, was studied to help elucidate how it provides a strong, adhesive glue-like function that anchors dragline silk. The purpose of our research was to express a portion of the PySp1 protein. PCR was used to amplify a segment of the PySp1 cDNA, which was then inserted into the pBAD-Thio/TOPO bacterial expression vector. E. coli was then transformed with the ligation mixture and colonies carrying the correct vector were identifies by restriction digestion and agarose gel electrophoresis. Transformants carrying the vector were induced with arabinose to express the PySp1 cDNA insert. Western blot analysis was then used to check for the expression of the PySp1 protein. With the results obtained, further experiments will be carries out in attempt to synthesize synthetic pyriform fibers