【作者】 奚丽丽；

【导师】 王兴智；

【作者基本信息】 东北师范大学 ， 细胞生物学， 2006， 硕士

【摘要】 作为一种安全、高效、快速的外源蛋白生产体系,植物病毒瞬时表达系统是近年发展起来的一种新型蛋白表达方式。它避免了转基因植物存在的外源基因通过有性生殖过程向环境扩散的危险,并且,由于宿主范围的限制,植物病毒载体不会对人或动物构成危害。利用该表达体系可大量生产外源蛋白,改善了转基因植物普遍存在的外源基因表达水平低的状况。该表达体系无需进行植物组织培养和阳性植株筛选,可以节省大量的时间和人力。尽管如此,只有很少的豆科病毒载体既可以用于基因表达又能用于基因沉默。之所以选择豆科植物作为实验材料是因为豆科植株中蛋白含量很高。本实验以绿色荧光蛋白(GFP)为报告基因,建立并优化了PEBV瞬时表达体系以提高豆科植物的商业价值。结果表明,本实验成功地建立并优化了以PEBV为载体的外源蛋白瞬时表达体系,报告基因gfp实现了在豆角中的系统扩散和大量表达。更多还原

【Abstract】 As a safe, efficient and rapid production system for foreign proteins plant virus transientexpression system is a novel strategy, which was developed in recent years. It avoids the riskof transgene flowing into the environment through sexual reproduction, which is a safetyconcern about stable transgenic plants. Based on the host range restriction, plant virus vectorwill not infect human beings or animals. The protein produced in this system achieves highpercentage of total soluble proteins, which is an improvement of the low-level expression instable transgenic plants. Moreover, the time and labor-consuming tissue culture and positiveselection steps can be avoided.Even with these advances, there are only a limited number of legume virus vectors thatare suitable both for the gene expression and for gene silencing in the legume species.Legume species are favorable as the experiment materials because of the high level ofproteins they contained. With GFP as the report gene, the transient expression systememploying Pea Early Browning Virus-based (PEBV) vector was established and optimized inorder to verify the possibility of using legume plants to increase the production of valuableproteins.The results suggested that the PEBV transient expression system has been establishedsuccessfully and the GFP gene has been spread systemically and expressed at high level inBean Pod.更多还原