Recombinant DNA production of spider silk proteins.
Bottom Line: Moreover, silks are biocompatible and biodegradable protein-based materials.Recent advances in genetic engineering make it possible to produce recombinant silks in heterologous hosts, opening up opportunities for large-scale production of recombinant silks for various biomedical and material science applications.We review the current strategies to produce recombinant spider silks.
Affiliation: Department of Biomedical Engineering, Tufts University, Medford, MA, 02155, USA.Show MeSH
Mentions: Unicellular organisms, such as bacteria and yeast, have been investigated as host systems for recombinant silks. A gram-negative, rod-shaped bacterium E. coli is a well-established host for industrial scale production of proteins. Therefore, the majority of recombinant spider silks have been produced in E. coli (Lewis et al., 2011; Fahnestock and Irwin, 1997; Wang et al., 2006; Rabotyagova et al., 2009; Rabotyagova et al., 2010; An et al., 2011; An et al., 2012; Teulé et al., 2012a). E. coli is easy to manipulate, has a short generation time, is relatively low cost and can be scaled up for larger amounts protein production. The recombinant DNA approach enables the production of recombinant spider silks with programmed sequences, secondary structures, architectures and precise molecular weight (Rabotyagova et al., 2011). There are four main steps in the process: (i) design and assembly of synthetic silk-like genes into genetic ‘cassettes’, (ii) insertion of this segment into a DNA vector, (iii) transformation of this recombinant DNA molecule into a host cell and (iv) expression and purification of the selected clones. Figure 2 summarizes the recombinant DNA approach used to prepare silk-like proteins.
Affiliation: Department of Biomedical Engineering, Tufts University, Medford, MA, 02155, USA.