Construction of spider silk functional platform via intein trans-splicing.

Author: Senzhu LIN ¹ ; Gefei CHEN ¹ ; Qing MENG ¹
Author Information

1. Institute of Biological Sciences and Biotechnology, Donghua University, Shanghai 201620, China.
Publication Type:Journal Article
Keywords: functionalized modification; recombinant aciniform silk protein; recombinant spider silk; small ubiquitin-related modifier; split intein
MeSH: Animals; Inteins; Protein Splicing; Recombinant Proteins; chemistry; Silk; chemistry; Small Ubiquitin-Related Modifier Proteins; chemistry; Spiders; Trans-Splicing
From: Chinese Journal of Biotechnology 2016;32(12):1704-1714
CountryChina
Language:Chinese
Abstract: To provide technical support for spider silk functional modification, we developed a simple and efficient functional platform via intein trans-splicing. Small ubiquitin-related modifier protein (SUMO) was fused to the recombinant spider silk protein (W2CT) by peptide bond via S0 split intein Ssp DnaB trans-splicing, resulting in a protein SUMOW2CT. However, incorporation of exogenous protein led to mechanical property defect and lower fiber yield, and also slowed down the fiber assembly velocity but no obvious differences in supercontraction and chemical resistance when compared with fibers from W2CT (W). SUMO protease digestion showed positive results on the fibers, indicating that the SUMO protein kept its native conformation and bioactive. Above all, this work provides a technical support for spider silk high simply and efficient functionalized modification.