Production and purification of recombinant DP1B silk-like protein in plants |
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Authors: | Barr Leslie A Fahnestock Stephen R Yang Jianjun |
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Institution: | (1) Central Research and Development, E. I. DuPont de Nemours & Co. Experimental Station, Wilmington, DE, 19880-0402 |
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Abstract: | Spider dragline silk of Nephila clavipes consists of two highly repetitive proteins, spidroin 1 and spidroin 2. To develop a plant platform for production of recombinant silk-like protein, two plant-optimized DP1B genes were synthesized to mimic spidroin 1. DP1B-8P encodes for a 64 kD DP1B silk-like protein and DP1B-16P for a 127 kD DP1B silk-like protein. Both genes have been introduced into Arabidopsis for leaf-based production driven by the 35S promoter and for seed-specific production driven by the -conglycinin ![agr](/content/n8g7505124471r72/xxlarge945.gif) subunit promoter, respectively. They have also been expressed in somatic soybean embryos under the control of the -conglycinin ![agr](/content/n8g7505124471r72/xxlarge945.gif) subunit promoter. The results demonstrate the synthesis and accumulation of DP1B silk-like protein in leaves and seeds of Arabidopsis, as well as in somatic soybean embryos. They suggest that seeds are the more preferred tissue for DP1B production since they offer higher production yield and quality. In addition, a simplified protocol for purifying DP1B from plant tissue is discussed. |
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Keywords: | Arabidopsis Heterologous protein expression Plant genetic engineering Spider dragline silk Silk-like protein Somatic soybean embryo |
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