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University of the Pacific Theses and Dissertations

2024

Bacteria

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Quantitative Shotgun Proteomic Analysis Of Bacteria After Overexpression Of Recombinant Spider Miniature Spidrion, Masp1, Kathryn P. Randene Jan 2024

Quantitative Shotgun Proteomic Analysis Of Bacteria After Overexpression Of Recombinant Spider Miniature Spidrion, Masp1, Kathryn P. Randene

University of the Pacific Theses and Dissertations

Spider silk has extraordinary mechanical properties, displaying high tensile strength, elasticity, and toughness. Given the high performance of natural fibers, one of the long-term goals of the silk community is to manufacture large-scale synthetic spider silk. This process requires vast quantities of recombinant proteins for wet-spinning applications. Attempts to synthesize large amounts of native size recombinant spidroins in diverse cell types have been unsuccessful. In these studies, we design and express recombinant miniature black widow (Latrodectus hesperus) MaSp1 spidroins in bacteria that incorporate the NTD and CTD, along with varying numbers of codon-optimized internal block repeats. Following spidroin overexpression, we …