| 초록 |
Dragline spider silk has a number of notable mechanical properties such as high tensile strength and elasticity. Strong natural fibers with good tensile and compressive properties would be useful for many applications in medicine, such as sutures, membranes and temporary scaffolds for tissue engineering. This silk protein is composed of two proteins, MaSP 1 and 2, whose partial cDNAs have been cloned and sequenced (Lewis et al. 1992). Recently, recombinant spider silks have been generated from synthetic spider silk genes using Escherichia coli as the heterologous host. While recombinant proteins have been successfully generated from these genetically engineered systems (Winkler et al. 1999 Winkler et al. 2000, Szela et al. 2000), a significant limitation in these studies has been the difficulties in high-level producing genetically engineered silk-like proteins (Lewis et al. 1992, S. Winkler et al. 2000). Presently the in vivo production of artificial polymeric proteins is just starting and yields are still low. Thus, a need to overcome the low expression is indispensable. In this study, we describe the efficient process development during high-cell density cultivation of recombinant E. coli for the production of recombinant silk protein. We also describe simple procedure for purification of silk protein. The methodology described will allow production of sufficient quantities of the protein for many potential uses including production of artificial synthetic fibers. |
