Characterization of two full-length Araneus ventricosus major ampullate silk protein genes

Abstract

Major ampullate silk is noted for its great tensile strength and extensibility. The impressive material properties of major ampullate silk result from their component proteins that encoded by members of the spidroin (spider fibroin) gene family. Although the major ampullate spidroin type has evolved multiple variants within specific-species, most sequences are fragmented. Here, we present two complete major ampullate spidroin genes from the orb-weaving spider Araneus ventricosus. Due to the abundant GPG motifs in their repetitive region, the two MaSp genes were grouped in MaSp2 subclass and named MaSp2C and MaSp2D, respectively. Analysis of the full-length gene sequences reveals that both of them include a single enormous exon (10,851 bp for MaSp2C and 8640 bp for MaSp2D) that mainly translates into a central repetitive region containing multiple amino acid motifs that can be organized into five ensemble types. We use gene-specific PCR primers to search the cDNA from major ampullate glands and find evidence for alternative splicing of MaSp2D transcripts into a minor spliceoform lacking the entire repetitive domain as well as the partial terminal regions. Our results not only provide new templates for protein-based materials with tailored properties, but suggest gene and transcriptional diversity of major ampullate silk.

Keywords: Alternative splicing; Araneus ventricosus; MaSp.