Expanding Canonical Spider Silk Properties Through A Dna Combinatorial Approach

The properties of native spider silk vary within and across species due to the presence of different genes containing conserved repetitive core domains encoding a variety of silk proteins. Previous studies seeking to understand the function and material properties of these domains focused primarily on the analysis of dragline silk proteins, MaSp1 and MaSp2. Our work seeks to broaden the mechanical properties of silk-based biomaterials by establishing two libraries containing genes from the repetitive core region of the nativeLatrodectus hesperussilk genome (Library A: genesmasp1,masp2,tusp1,acsp1; Library B: genesacsp1,pysp1,misp1,flag). The expressed and purified proteins were analyzed through Fourier Transform Infrared Spectrometry (FTIR). Some of these new proteins revealed a higher portion of beta-sheet content in recombinant proteins produced from gene constructs containing a combination ofmasp1/masp2andacsp1/tusp1genes than recombinant proteins which consisted solely of dragline silk genes (Library A). A higher portion of beta-turn and random coil content was identified in recombinant proteins frompysp1andflaggenes (Library B). Mechanical characterization of selected proteins purified from Library A and Library B formed into films was assessed by Atomic Force Microscopy (AFM) and suggested Library A recombinant proteins had higher elastic moduli when compared to Library B recombinant proteins. Both libraries had higher elastic moduli when compared to native spider silk proteins. The preliminary approach demonstrated here suggests that repetitive core regions of the aforementioned genes can be used as building blocks for new silk-based biomaterials with varying mechanical properties.