Agriculture and AgriFood Canada, Southern Crop Protection and Food Research Center, 1391 Sandford Street, London, Ont., Canada, N5V 4T3.
Plant Biotechnol J. 2004 Sep;2(5):431-8. doi: 10.1111/j.1467-7652.2004.00087.x.
Spider dragline silk is a unique biomaterial and represents nature's strongest known fibre. As it is almost as strong as many commercial synthetic fibres, it is suitable for use in many industrial and medical applications. The prerequisite for such a widespread use is the cost-effective production in sufficient quantities for commercial fibre manufacturing. Agricultural biotechnology and the production of recombinant dragline silk proteins in transgenic plants offer the potential for low-cost, large-scale production. The purpose of this work was to examine the feasibility of producing the two protein components of dragline silk (MaSp1 and MaSp2) from Nephila clavipes in transgenic tobacco. Two different promoters, the enhanced CaMV 35S promoter (Kay et al., 1987) and a new tobacco cryptic constitutive promoter, tCUP (Foster et al., 1999) were used, in conjunction with a plant secretory signal (PR1b), a translational enhancer (alfalfa mosaic virus, AMV) and an endoplasmic reticulum (ER) retention signal (KDEL), to express the MaSp1 and MaSp2 genes in the leaves of transgenic plants. Both genes expressed successfully and recombinant protein accumulated in transgenic plants grown in both greenhouse and field trials.
蜘蛛牵引丝是一种独特的生物材料,代表了自然界已知的最强纤维。由于它几乎和许多商业合成纤维一样强韧,因此适用于许多工业和医疗应用。这种广泛应用的前提是能够以具有成本效益的方式大量生产,以满足商业纤维制造的需求。农业生物技术和在转基因植物中生产重组牵引丝蛋白为低成本、大规模生产提供了可能性。本工作旨在研究利用 Nephila clavipes 在转基因烟草中生产牵引丝的两种蛋白成分(MaSp1 和 MaSp2)的可行性。我们使用了两种不同的启动子,增强型 CaMV 35S 启动子(Kay 等人,1987 年)和新的烟草隐蔽组成型启动子 tCUP(Foster 等人,1999 年),结合植物分泌信号(PR1b)、翻译增强子(苜蓿花叶病毒,AMV)和内质网(ER)保留信号(KDEL),在转基因植物的叶片中表达 MaSp1 和 MaSp2 基因。两个基因都成功表达,重组蛋白在温室和田间试验中生长的转基因植物中积累。