大鼠IP-10在大肠杆菌中的表达及其生物学活性的研究
发布时间：2019-03-24 10:50
【摘要】：目的： 趋化因子(chemokine)在机体的免疫反应和疾病的发生发展过程中起着重要的作用。它们在炎性条件下由单核细胞、内皮细胞及淋巴细胞表达，参与免疫活性细胞的活化和移行，从而调节免疫应答。近年来发现趋化因子与中枢神经系统(central nervous system，CNS)炎症有关，如多发性硬化(mutiple sclerosis，MS)、实验性变态反应性脑脊髓炎(experimental allergic encephalomyelitis，EAE)。CNS炎症性疾病的基本病理过程是炎性细胞募集浸润，而趋化因子在调节淋巴细胞进入CNS组织的过程中发挥着极其重要的作用，其中γ-干扰素诱导蛋白10(interferon-γ-inducible protein-10，IP-10)在CNS炎性反应过程中的作用颇受关注。IP-10属于趋化因子超家族CXC类ELR-亚族，其主要作用是对多种炎症细胞包括T淋巴细胞、单核细胞和自然杀伤细胞具有趋化活性。许多研究表明当CNS发生感染时，小胶质细胞开始活化并且大量表达IP-10，IP-10介导炎性细胞浸润，从而启动或促进CNS的炎性反应进程。为进一步研究IP-10的生物学活性及其在CNS炎症性疾病中的作用，本研究构建了大鼠pET-32a(+)-IP10原核表达质粒，并转入大肠杆菌BL21(DE3)中表达，通过亲和层析纯化出IP-10融合蛋白，对其活性和趋化功能作初步的鉴定，，也为探寻以IP-10为靶治疗CNS炎症性疾病的治疗新措施奠定了基础。 方法： 本研究由两部分组成：第一部分为大鼠IP-10原核表达质粒的构建。首先从GenBank上获取大鼠IP-10 cDNA基因序列，使用引物设计软件Primer Premier 5.0进行引物设计，从中选择最佳引物。考虑到后续的纯
[Abstract]:Objective: chemokine (chemokine) plays an important role in immune response and disease development. They are expressed by monocytes, endothelial cells and lymphocytes under inflammatory conditions, and participate in the activation and migration of immunoactive cells, thus regulating the immune response. In recent years, chemokine has been found to be associated with (central nervous system,CNS inflammation in the central nervous system, such as multiple sclerosis (mutiple sclerosis,MS) and experimental allergic encephalomyelitis (experimental allergic encephalomyelitis,). (EAE). The basic pathological process of inflammatory diseases in CNS is inflammatory cell recruitment and infiltration, and chemokine plays an important role in regulating lymphocyte entry into CNS tissue. The role of interferon-纬-induced protein 10 (interferon- 纬-inducible protein-10,IP-10) in the inflammatory process of CNS has attracted much attention. IP-10 belongs to the chemokine superfamily CXC-like ELR-subfamily. Its main function is chemotactic activity to a variety of inflammatory cells, including T lymphocytes, monocytes and natural killer cells. Many studies have shown that when CNS infection occurs, microglia begin to activate and express a large number of IP-10,IP-10-mediated inflammatory cell infiltration, thus initiating or promoting the inflammatory process of CNS. In order to further study the biological activity of IP-10 and its role in inflammatory diseases of CNS, the prokaryotic expression plasmid of rat pET-32a ()-IP10 was constructed and transformed into E. coli BL21 (DE3). The IP-10 fusion protein was purified by affinity chromatography, and its activity and chemotactic function were preliminarily identified. It also laid a foundation for exploring new therapeutic measures for treating inflammatory diseases of CNS with IP-10 as target. Methods: this study consists of two parts: the first part is the construction of prokaryotic expression plasmid of rat IP-10. At first, the sequence of rat IP-10 cDNA gene was obtained from GenBank. Primer design software Primer Premier 5.0 was used to design the primer, and the best primer was selected from the primer design software Primer Premier 5.0. Considering the subsequent purity,
【学位授予单位】：四川大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R346
本文编号：2446253
[Abstract]:Objective: chemokine (chemokine) plays an important role in immune response and disease development. They are expressed by monocytes, endothelial cells and lymphocytes under inflammatory conditions, and participate in the activation and migration of immunoactive cells, thus regulating the immune response. In recent years, chemokine has been found to be associated with (central nervous system,CNS inflammation in the central nervous system, such as multiple sclerosis (mutiple sclerosis,MS) and experimental allergic encephalomyelitis (experimental allergic encephalomyelitis,). (EAE). The basic pathological process of inflammatory diseases in CNS is inflammatory cell recruitment and infiltration, and chemokine plays an important role in regulating lymphocyte entry into CNS tissue. The role of interferon-纬-induced protein 10 (interferon- 纬-inducible protein-10,IP-10) in the inflammatory process of CNS has attracted much attention. IP-10 belongs to the chemokine superfamily CXC-like ELR-subfamily. Its main function is chemotactic activity to a variety of inflammatory cells, including T lymphocytes, monocytes and natural killer cells. Many studies have shown that when CNS infection occurs, microglia begin to activate and express a large number of IP-10,IP-10-mediated inflammatory cell infiltration, thus initiating or promoting the inflammatory process of CNS. In order to further study the biological activity of IP-10 and its role in inflammatory diseases of CNS, the prokaryotic expression plasmid of rat pET-32a ()-IP10 was constructed and transformed into E. coli BL21 (DE3). The IP-10 fusion protein was purified by affinity chromatography, and its activity and chemotactic function were preliminarily identified. It also laid a foundation for exploring new therapeutic measures for treating inflammatory diseases of CNS with IP-10 as target. Methods: this study consists of two parts: the first part is the construction of prokaryotic expression plasmid of rat IP-10. At first, the sequence of rat IP-10 cDNA gene was obtained from GenBank. Primer design software Primer Premier 5.0 was used to design the primer, and the best primer was selected from the primer design software Primer Premier 5.0. Considering the subsequent purity,
【学位授予单位】：四川大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R346
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本文编号：2446253
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