Biomimetic manufacturing of customised novel fibre proteins for specialised applications

• Autorzy: Teulé F. , Aubé C. , Ellison M. , Abbott A.
• Języki publikacji: EN

Abstrakty

EN

Using spider silk and collagen as a model, we have investigated the role that various protein primary structural components play in fibre production. Spidroins, spider dragline silk protein components, are essentially characterised by an amino-acid repeat containing a glycine-rich motif (amorphous) followed by an alanine-rich motif (crystalline, putatively responsible for fibre strength). We have tested the importance of alanine runs in these proteins and the role of this motif in the mechanical properties of the resulting fibre. To test the importance of alanine-rich motifs in the spidroin-1 proteins, we engineered three types of spidroin-1-like genes containing sequence encoding for different amounts of alanine repeats in the protein (normal, low, and no alanine residues). We also have engineered three copolymer collagen-spidroin-1 genes using each of the three spidroin-1 synthetic genes. These copolymers were mimicked on the existing natural block collagen-silk-like protein copolymer found in the byssus thread of marine mussels. All of these constructs were introduced in yeast (Pichia pastoris) for protein production. We are currently purifying each of the recombinant proteins for structural analysis (CD-spectroscopy).

Słowa kluczowe

EN

spider silk; spidroin; collagen; synthetic genes; Pichia pastoris

PL

jedwab; kolagen; geny syntetyczne; Pichia pastoris

• Wydawca: Lodz University of Technology, Faculty of Material Technologies and Textile Design
• Czasopismo: Autex Research Journal
• Rocznik: 2003
• Tom: Vol. 3, no. 4
• Strony: 160--165
• Opis fizyczny: Bibliogr. 10 poz.

Twórcy

autor

Teulé F.

• Dept. of Genetics and Biochemistry, Clemson University, Clemson, SC 29634, USA

autor

Aubé C.

• Dept. of Genetics and Biochemistry, Clemson University, Clemson, SC 29634, USA

autor

Ellison M.

• School of Materials Science and Engineering, Clemson University, Clemson, SC 29634, USA

autor

Abbott A.

• Dept. of Genetics and Biochemistry, Clemson University, Clemson, SC 29634, USA

Bibliografia

• 1. Gosline, J., Guerette, P., Ortlepp., C., Savage, K. (1999) The mechanical design of spider silks: From fibroin sequence to mechanical function. J. Exp. Biol. 202, pp. 32295-3303
• 2. Xu, M., and Lewis, R.V. (1990) Structure of a protein superfibre: spider dragline silk. Proc. Natl. Acad. Sci. USA, 87, pp. 7120-7124
• 3. Hinman, M.B., and Lewis, R.V. (1992) Isolation of a clone encoding a second dragline silk fibroin. J. Biol. Chem., 267 (27), pp. 19320-19324
• 4. Hayashi, C.Y., Shipley, N.H., and Lewis, R.V. (1999) Hypotheses that correlate the sequence, structure, and mechanical properties of spider silk proteins. Int. J. of Biol. Macromolecules, 24, pp. 271-275
• 5. Heslot, H. (1998) Artificial fibrous proteins: A review. Biochimie, 80, pp. 19-31
• 6. Lazaris, A., Arcidiacono, S., Huang, Y., Zhou, J.F., Duguay, F., Chretien, N., Welsh, E.A., Soares, J.W., and Karatzas, C.N. (2002) Spider silk fibres spun from soluble recombinant silk produced in mammalian cells. Science, 295, pp. 472-476
• 7. Scheller, J., Guhrs, K.H., Grosse, F., and Conrad, U. (2001) Production of spider silk proteins in tobacco and potato. Nature, 19, pp. 573-577
• 8. Wang, T., Deom, C.M., and Hussey, R.S. (1998) Identification of a Meloidogyne incognita cuticle collagen gene and characterisation of the developmental expression of three collagen genes in parasitic stages. Mol. Biol. Parasitol., 93, pp. 131-134
• 9. Sambrook J., Fritsch E.F., and Maniatis T.E. (1989) Molecular cloning A laboratory manual. Second Edition; Cold Spring Harbor Laboratory
• 10. Birnboim H.C., Doly J. (1979) A rapid alkaline extraction procedure for screening recombinant DNA. Nucleic Acids Research, 7,pp. 1513-1518