【作者】 扈廷茂；

【导师】 张学明；

【作者基本信息】 内蒙古大学 ， 微生物学， 2006， 硕士

【摘要】 人内皮抑素(Endostatin)是第一个进入临床研究的内源性血管生成抑制剂，是胶原蛋白ⅩⅧC末端非胶原区NC1结构域的20KD的片段。它可以抑制内皮细胞的增殖和转移，从而降低肿瘤内血管生成，阻断营养通路，达到杀死肿瘤细胞的目的。其最大的优点是无毒性和无抗药性。植物生物反应器生产药物蛋白成本低，生产等量目的蛋白只需要花其它表达系统所需费用的10％，而且一旦建立起稳定而高效的表达系统，只需扩大种植面积就可大量生产目的蛋白。本研究将人内皮抑素基因重组于植物双元表达载体pGA643，得到重组质粒pGE；用三亲融合法将其导入农杆菌LBA4404中，采用叶盘法转化烟草，经诱导与筛选，获得了卡那霉素抗性植株；提取抗性植株总DNA，通过PCR、斑点杂交和Southern杂交检测，筛选出了整合有外源基因的转化植株，为进一步研究人内皮抑素在烟草中的表达及生物学功能奠定了基础。更多还原

【Abstract】 Endostatin, a carboxy-terminal 20KDa fragment of the non-collagenous (NC) 1 domain of collagen Ⅹ Ⅷ, is the first endogenous angiogenesis inhibitor entering human clinical therapy. It can inhibit endothelial cell migration and multiplication to reduce the vascularization of tumors and finally kill tumors. The great advantage of the agent lies in its less toxicity and not inducing drug resistance when used. The use of plant bioreactors for the production of recombinant pharmaceutical proteins is safe and inexpensive. Cultivating transgenic plants on an agricultural scale will produce almost unlimited supplies of recombinant proteins for uses in medicine. The plant expression vector pGE, carrying a human endostatin gene, was constructed. pGE was introduced into Agrobacterium tumefaciens by triparental mating. Tobacco leaf discs were transformed via Agrobacterium Mmefaciens mediated procedure. Transgenic tobacco plantlets grew in induction medium and plants resistant to kanamycin were obtained. PCR, Dot blotting and Southern hybridization analysis confirmed that human endostatin gene had integrated into the genome of tobacco.更多还原