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