CAJ | 학술논문

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.
골질소송증엄중위협착인류적건강,해어강개소시일충치료골질소송적유효약,단시앙귀적개격한제료타적사용.본연구이생채작위생물반응기래표체강개소중생물활성교고적해어강개소,이기통과선식표체강개소적생채체도방지혹치료골질소송목적,동시가대대강저생산성본.수선안식물편애밀마자설계합성료편마해어강개소적msCT기인,기차고필도전기인산품안전성문제,구건식물쌍원표체재체p35S-2300-twinT-DNA::pil-msCT::noster,이용농간균개도진행생채전화.경PCR화Southern blot학인득도료12주독립적전기인식주.동시RT-PCR분석표명해어강개소재To대전기인식주중성공표체,이차표체량존재현저차이,위이용식물생물반응기생산치료용강개소전정료기출.
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.