甲型H1N1流感病毒重要抗原HA1、NAe蛋白的表达及应用内科学(传染病)专业论文.docxVIP

甲型 甲型 H1N1 流感病毒重要抗原 HA1、NAe 蛋白的表达及应用 广州医 广州医科大学硕士学位论文 PAGE PAGE 13 PAGE PAGE 10 化学发光条带，而正常人检测无发光条带显示，表明表达蛋白具有较好的抗原 活性。 第二部分 1) 在 Western blot 检测中，纯化前后的重组蛋白均可与甲流患者血清的抗体结合 产生明显的特异性免疫反应条带； 2) 34 例临床标本中，表达蛋白检测显示，IgG 抗体阳性 28 例，阴性 6 例；IgM 抗体阳性 1 例，阴性 33 例； 3) 抗流感病毒 A 型抗体 IgM 检测试剂盒检测显示 IgM 抗体可疑阳性 1 例，阴性 33 例。 结论 甲型H1N1流感病毒HA和NA基因去除了信号肽和疏水区后可在原核表达系统 中高效表达，成功表达的甲型H1N1流感病毒重要抗原HA1、NAe蛋白具有免疫活 性，可用于Western blot技术检测甲流IgG、IgM抗体。 关键词 甲型 H1N1 流感病毒；血凝素；神经氨酸酶；原核表达；甲型流感病 毒 IgM 抗体 Application and expression of the antigenic protein HA1 and NAe from influenza A-H1N1 virus Major: Internal Medicine (Infectious Disease ) M.Candidate: Jianbo Zhong Supervision: Prof. Shaojing Wei Abstract Recent years, it is an important way to diagnose the patients with infection of influenza A virus that serum antibodies are tested with the engineering viral antigens. Although a lot of studies have been made for the engineering antigens of Hemagglutinin(HA) and Neuraminidase(NA), internationally and nationally, some problems are needed to be solved, such as which is best system for expression of engineering antigens of HA and NA, how to get expression products with good quality and quantity and the impact factors. There are several of expression system for the engineering viral antigens, mainly including eukaryotic expression system, cell free expression system and prokaryotic expression system. The products of eukaryotic expression system are easier to be purified and protein activity is not affected. However, the product quantity is lower and the cost is higher. In addition, the cell culture is time-consumer and not easier to be cultivated. Cell-free expression technology is not mature enough to be generally applied. For prokaryotic expression system, the product quantity is higher, easier to handle, saving time, and usually used for a large scale of protein expression. The expressed products existing in inclusions, as insoluble particles, could be purified by precipitation. This expression system