抗PDI因子(Anti-PDI Factor)生物检测系统的构建
时间:2021-03-04 21:07 来源:毕业论文 作者:毕业论文 点击:次
摘要卵巢癌是目前死亡率最高的妇科疾病,70%的患者因在晚期确诊而难以得到有效的治疗。现在需要从天然或者合成的化合物中筛选一种强有力的、具有选择性的、安全的口服抗PDI因子,回复肿瘤细胞的药敏性,用于卵巢癌的伴侣治疗。本研究根据文献me-too合成一种抗PDI因子,用于本课题构建的抗PDI因子检测系统的验证。针对抗PDI因子细菌检测体系,本研究采用基因敲除、基因置换等技术构建大肠杆菌dsbA突变体,然后需要采用构建报告基因等手段来构建,最终通过细菌报告基因表达来指示待测物质的抗PDI活性。抗PDI因子的me-too合成和大肠杆菌dsbA突变体的成功构建为最终抗PDI因子检测系统的构建打下了坚实的基础。63953 毕业论文关键词 卵巢癌 抗PDI因子 检测系统 突变体 毕业设计说明书(论文)外文摘要 Title :The Construction of an anti-PDI factor testing system in Escherichia coli dsbA mutant Abstract Ovarian cancer is one of the leading causes of death in women with gynecological cancers in the world. About 70% of ovarian cancer cases are diagnose data late stage and therefore, poorly treatable. With the resistant tumoral cells being appeared, it is necessary to R&D one potent, target, and safe small-molecule inhibitors of PDI from natural or synthetic compounds to recover the sensitivity of these anti-tumor drugs. In the present study, an anti-PDI factor is chemically synthesized me-too to verify the availability of anti-PDI screening system. As for the construction of anti-PDI screening system, some interdisciplinary methods were used here, such as gene deletion, gene substitution and finally construct target report gene in Escherichia coli dsbA mutant. The synthesis of anti-PDI factor and the construction of E.coli dsbA mutant are basic to test anti-PDI factor detection system next. Keywords ovarian cancer anti-PDI factor detection system mutant 目 次 1.绪论 1 1.1卵巢癌的简介 1 1.2卵巢癌的治疗手段 1 1.3病原细胞的抗药 2 1.4人体内的PDI 3 1.5大肠杆菌内的dsbA基因 3 1.6博来霉素抗性基因ble 4 1.7研究工作的发展和应用前景 4 2.实验 5 2.1实验目的 5 2.2实验原理 5 2.3实验仪器和材料 6 2.4实验涉及的方法和技术 7 2.4.1 Ugi反应 7 2.4.2 PCR 10 2.4.3细菌转化 10 2.4.4凝胶电泳 10 2.4.5基因敲除 11 2.5实验操作和结果 11 2.5.1经典Anti-PDI因子的me-tooaa化学合成 11 2.5.2大肠杆菌中与人体PDI相对应的dsbA基因的敲除 13 3.结果与讨论 31 3.1化合物1的1H NMR谱图解析 31 3.2酶连反应载体选择的重要性 31 3.3通过正交实验获取提取质粒pDGICZ的最佳条件 (责任编辑:qin) |