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Molecular design and production of silk-like materials with cell adhesion activity in Echerichia coli

時(shí)間:2004-12-07
關(guān)鍵詞:Molecular design production silk materials cell adhesion activity Echerichia coli 來源：International Symposium on Biological Polyesters ,Auguest 22-27, 2004

 

J. Yao1, and T. Asakura2

 

1 The Key Laboratory of Advanced Textile Materials and Manufacturing Technology of Ministry of Education, Collage of Materials and Textile, Zhejiang University of Sciences, Hangzhou 310018, China

2 Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan.

Introduction: Silks have many inherent superior properties as biomaterials in the viewpoints of mechanical properties, environmental stability and biocompatibility.  In this research, silk-like materials were designed and produced with genetically engineering synthesis method, which were generally combined with the structural units selected from Bombyx mori silk fibroin and animal elastin, and the functional units, e.g., RGD sequence, derived from fibronectin in order to improve the cell adhesion property. 

Materials and Methods: Bacterial growth in rich medium, DNA manipulations, and transformation conditions were performed as described previously.1,2  Batch cultures were performed on MDL-6C Fermentor (B. E. Marubishi Co.), with 1.2 L working volume. 

 

                                     

                                      Fig. 1  Cell adhesion activity of silk-like materials

Results and Discussion: The synthetic genes were constructed in Escherichia coli and the fusion proteins were expressed, purified and characterized. 13C CP/MAS NMR data suggested that the conformation of silk-like materials could be easily controlled by the treatment with different solvents.  Moreover, the silk-like materials showed higher cell adhesive and growth activities than collagen due to the incorporation of RGD unit, suggesting that these materials could be the candidates for the medical applications after suitable processing.

 

[1]     Sambrook, J., et al., 1989. Molecular Cloning. A Laboratory Manual, Cold Spring Harbor Press, Cold Spring Harbor, NY. [2]      Yao, J. and Asakura, T., 2003. J. Biochem. 133, 147-154.

       論文來源：International Symposium on Biological Polyesters ,Auguest 22-27, 2004