【摘要】 骨质疏松症严重威胁着人类的健康,鲑鱼降钙素是一种治疗骨质疏松的有效药,但是昂贵的价格限制了它的使用。本研究以生菜作为生物反应器来表达降钙素中生物活性较高的鲑鱼降钙素,以期通过鲜食表达降钙素的生菜达到防止或治疗骨质疏松目的,同时可大大降低生产成本。首先按植物偏爱密码子设计合成了编码鲑鱼降钙素的msCT基因,其次考虑到转基因产品安全性问题,构建植物双元表达载体p35S-2300-twinT-DNA::pil-msCT::noster,利用农杆菌介导进行生菜转化。经PCR和Southern blot确认得到了12株独立的转基因植株。同时RT-PCR分析表明鲑鱼降钙素在To代转基因植株中成功表达,而且表达量存在显著差异,为利用植物生物反应器生产治疗用降钙素奠定了基础。更多还原

【Abstract】 Osteoporosis (OP) threatens human health seriously. Salmon calcitonin (sCT) has been proven to be an effective, but expensive drug for curing this disease. In order to obtain safe and cheap sCT and explore a new alternative method for preventing and curing OP, lettuce (Lactuca sativa cv. Zhouye Lettuce)was explored as bioreactor to produce sCT. According to plant codons usage bias, mutated sCT (msCT) gene was designed and linked to the 3’-end of pilus protein gene (pil) resulting in a fusion gene pil-msCT. The plant binary vector p35S-2300-twinT-DNA::pil-msCT::noster was constructed and introduced into lettuce plants via Agrobacterium tumefaciens -mediated method. In total, 12 independent transgenic lettuce plants were confirmed by PCR and Southern blot analysis. The expression of msCT in transgenic To plants was analyzed by RT-PCR. The results proved that the salmon calcitonin mutant has been expressed successfully in lettuce plants at transcriptional level, which will lay a foundation for using plant as bioreactor to produce effective therapeutic calcitonin.更多还原