约氏疟原虫子孢子侵入相关基因分析及235kDa棒状体蛋白研究
发布时间：2018-09-04 11:19
【摘要】： 蚊媒传播的疟疾是一种严重危害人类健康的热带传染病,在全世界人群中具有很高的发病率和致死率。近年来疟原虫多药抗性株的出现与迅速扩散,以及蚊媒对杀虫剂耐受性的增加,目前又无有效的抗疟疫苗,给疟疾防治工作带来很大的困难。传统方法已难于控制该病的流行,迫切需要为疟疾的防治工作提供新的方法和手段。疟疾是由蚊虫叮咬而传播,疟原虫子孢子一旦进入血循环,数分钟后便随血流跨过肝血窦枯否氏细胞(Kuppfer cell,KC)后主动粘附、迅速侵入肝细胞,并进行红外期增殖,然后进入红细胞,导致疟疾的发作。子孢子侵入肝细胞是疟疾感染的关键,而阻断虫体的侵入,即可防止疟疾感染,红外期阻断疫苗旨在阻断子孢子侵入宿主肝细胞,防止疟原虫红内期的发育,是疟疾疫苗的重要组成部分。 唾液腺子孢子感染性受发育调节。正常情况下,中肠内卵囊成熟后破裂,子孢子逸出,经血淋巴移行到唾液腺后2d,发育成为具有感染性的子孢子。因此研究疟原虫具有侵入肝细胞能力的唾液腺子孢子与不具侵入肝细胞能力的卵囊子孢子之间的差异分子具有重要意义。通过对疟原虫子孢子不同时相点侵入相关基因和蛋白的研究与鉴定,将有助于认识疟原虫生长发育的分子机制,可为疟疾的防治提供分子理论依据,提供药物和疫苗新的靶目标,为采取防治策略战胜疟疾打下了坚实的基础。 在后基因组时代,对基因功能的分析已成为当前的主要任务。而功能基因组学最大的技术挑战是关于蛋白表达的分析(蛋白质组学)。在本实验室前期的研究中,应用蛋白质组学的二维凝胶电泳(two-dimensional gel electrophoresis, 2-DE)技术、基质辅助激光解析电离飞行时间质谱(matrix-assisted laser desorption/inoization time of flying mass spectrometry, MALDI- TOF-MS)分析技术和蛋白质信息学等方法对约氏疟原虫卵囊子孢子和唾液腺子孢子这两个不同发育时期虫体的可溶性蛋白进行了分析研究。结果显示相对于卵囊子孢子,在唾液腺子孢子中确定比较明显的差异蛋白点有六个,分别为PyDp1~6(Plasmodium yoelii differential protein 1~6)。这些差异蛋白点主要为一些膜蛋白、表面蛋白和运动相关的蛋白。推测这些蛋白可能与疟原虫子孢子黏附、侵入肝细胞相关,可能为新的阻断策略和疟疾多价亚单位疫苗的靶目标。因此,加强这些差异表达蛋白的
[Abstract]:Mosquito-borne malaria is a tropical infectious disease that is seriously harmful to human health. It has a high incidence and mortality among people all over the world. In recent years, the emergence and rapid spread of multidrug resistant strains of Plasmodium and the increase of mosquito vector resistance to insecticides, but there is no effective antimalarial vaccine, has brought great difficulties to malaria control. Traditional methods have been difficult to control the epidemic of the disease, and it is urgent to provide new methods and means for malaria control. Malaria is transmitted by mosquito bites. Once the plasmodium sporozoite enters the blood cycle, it adheres actively with the blood flow across the hepatic sinusoidal Kupffer cells (Kuppfer cell,KC), rapidly invades the hepatocytes, proliferates in the infrared phase, and then enters the red blood cells. Lead to an attack of malaria. The invasion of sporozoite into hepatocytes is the key to malaria infection, and blocking the invasion of parasites can prevent malaria infection. The infrared phase blocking vaccine is designed to block the invasion of sporozoites into the host hepatocytes and prevent the development of the erythroid phase of Plasmodium falciparum. It is an important part of malaria vaccine. Salivary gland sporozoite infection is regulated by development. Under normal conditions, the oocysts in the midgut rupture after maturation, the sporozoites escape, and develop into infective sporozoites 2 days after the hemolymph migration to the salivary glands. Therefore, it is important to study the differential molecules between salivary gland sporozoites and oocystis that have the ability to invade the liver cells of Plasmodium falciparum. The study and identification of the genes and proteins involved in the invasion of Plasmodium sporozoites at different time points will help to understand the molecular mechanism of the growth and development of Plasmodium and provide molecular theoretical basis for malaria control. Providing new targets for drugs and vaccines lays a solid foundation for strategies to combat malaria. In the post-genome era, the analysis of gene function has become the main task. The biggest technical challenge in functional genomics is the analysis of protein expression (proteomics). Two dimensional gel electrophoresis (two-dimensional gel electrophoresis, 2-DE) technique of proteomics was used in the previous research in our laboratory. Matrix assisted laser desorption ionization time of flight mass spectrometry (matrix-assisted laser desorption/inoization time of flying mass spectrometry, MALDI- TOF-MS) and protein informatics for the analysis of plasmodium yoelii oocystis and salivary gland sporozoites at different developmental stages The soluble protein was analyzed and studied. The results showed that there were six distinct differentially expressed protein spots in salivary gland sporozoites compared with oocystid spores, which were called PyDp1~6 (Plasmodium yoelii differential protein 1 (6). These differential protein spots are mainly membrane proteins, surface proteins and exercise-related proteins. These proteins may be associated with plasmodium sporozoite adhesion and invasion of hepatocytes, and may be the target of new blocking strategies and multivalent subunit malaria vaccine. Therefore, the expression of these differentially expressed proteins
【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R383
本文编号：2221933
[Abstract]:Mosquito-borne malaria is a tropical infectious disease that is seriously harmful to human health. It has a high incidence and mortality among people all over the world. In recent years, the emergence and rapid spread of multidrug resistant strains of Plasmodium and the increase of mosquito vector resistance to insecticides, but there is no effective antimalarial vaccine, has brought great difficulties to malaria control. Traditional methods have been difficult to control the epidemic of the disease, and it is urgent to provide new methods and means for malaria control. Malaria is transmitted by mosquito bites. Once the plasmodium sporozoite enters the blood cycle, it adheres actively with the blood flow across the hepatic sinusoidal Kupffer cells (Kuppfer cell,KC), rapidly invades the hepatocytes, proliferates in the infrared phase, and then enters the red blood cells. Lead to an attack of malaria. The invasion of sporozoite into hepatocytes is the key to malaria infection, and blocking the invasion of parasites can prevent malaria infection. The infrared phase blocking vaccine is designed to block the invasion of sporozoites into the host hepatocytes and prevent the development of the erythroid phase of Plasmodium falciparum. It is an important part of malaria vaccine. Salivary gland sporozoite infection is regulated by development. Under normal conditions, the oocysts in the midgut rupture after maturation, the sporozoites escape, and develop into infective sporozoites 2 days after the hemolymph migration to the salivary glands. Therefore, it is important to study the differential molecules between salivary gland sporozoites and oocystis that have the ability to invade the liver cells of Plasmodium falciparum. The study and identification of the genes and proteins involved in the invasion of Plasmodium sporozoites at different time points will help to understand the molecular mechanism of the growth and development of Plasmodium and provide molecular theoretical basis for malaria control. Providing new targets for drugs and vaccines lays a solid foundation for strategies to combat malaria. In the post-genome era, the analysis of gene function has become the main task. The biggest technical challenge in functional genomics is the analysis of protein expression (proteomics). Two dimensional gel electrophoresis (two-dimensional gel electrophoresis, 2-DE) technique of proteomics was used in the previous research in our laboratory. Matrix assisted laser desorption ionization time of flight mass spectrometry (matrix-assisted laser desorption/inoization time of flying mass spectrometry, MALDI- TOF-MS) and protein informatics for the analysis of plasmodium yoelii oocystis and salivary gland sporozoites at different developmental stages The soluble protein was analyzed and studied. The results showed that there were six distinct differentially expressed protein spots in salivary gland sporozoites compared with oocystid spores, which were called PyDp1~6 (Plasmodium yoelii differential protein 1 (6). These differential protein spots are mainly membrane proteins, surface proteins and exercise-related proteins. These proteins may be associated with plasmodium sporozoite adhesion and invasion of hepatocytes, and may be the target of new blocking strategies and multivalent subunit malaria vaccine. Therefore, the expression of these differentially expressed proteins
【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R383
【参考文献】
相关期刊论文 前2条
1 许颖,张锡林,段建华,黄复生;改进的一维二维电泳法分析疟原虫子孢子蛋白质的差异表达[J];第三军医大学学报;2005年05期
2 许颖,张锡林,段建华,黄复生;约氏疟原虫卵囊、唾液腺子孢子差异表达蛋白的二维电泳-质谱分析[J];中国寄生虫病防治杂志;2005年03期
，本文编号：2221933
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