Abstract
Natural spider silk is one of the world’s toughest proteinaceous materials, yet a truly biomimetic spider silk is elusive even after several decades of intense focus. In this study, Next-Generation Sequencing was utilised to produce transcriptomes of the major ampullate gland of two Australian golden orb-weavers, Nephila plumipes and Nephila pilipes, in order to identify highly expressed predicted proteins that may co-factor in the construction of the final polymer. Furthermore, proteomics was performed by liquid chromatography tandem-mass spectroscopy to analyse the natural solid silk fibre of each species to confirm highly expressed predicted proteins within the silk gland are present in the final silk product. We assembled the silk gland transcriptomes of N. plumipes and N. pilipes into 69,812 and 70,123 contigs, respectively. Gene expression analysis revealed that silk gene sequences were among the most highly expressed and we were able to procure silk sequences from both species in excess of 1,300 amino acids. However, some of the genes with the highest expression values were not able to be identified from our proteomic analysis. Proteome analysis of “reeled” silk fibres of N. plumipes and N. pilipes revealed 29 and 18 proteins, respectively, most of which were identified as silk fibre proteins. This study is the first silk gland specific transcriptome and proteome analysis for these species and will assist in the future development of a biomimetic spider silk.
Highlights
Spider silk is an outstanding proteinaceous fibre that outperforms other natural and synthetic fibres in tensile strength analyses
Orb-weaving spiders have up to seven different glands associated with producing silks and glues used for web architecture [major ampullate gland (MA), minor ampullate (Mi), flagelliform (Flag), aggregate (Ag), and pyriform (Py) glands], protection of eggs [tubuliform glands (Tu)], and prey wrapping [aciniform glands (Ac)] [4]
We report on a silk-gland specific transcriptome analysis for these golden orb-weaving species, N. plumipes and N. pilipes
Summary
Spider silk is an outstanding proteinaceous fibre that outperforms other natural and synthetic fibres in tensile strength analyses. In combination with their genome assembly for N. clavipes, Babb et al characterised spidroin expression within the distinct silk glands [27]. We report on a silk-gland specific transcriptome analysis for these golden orb-weaving species, N. plumipes and N. pilipes.
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