約氏瘧原蟲子孢子侵入相關(guān)基因分析及235kDa棒狀體蛋白研究
發(fā)布時間：2018-09-04 11:19
【摘要】： 蚊媒傳播的瘧疾是一種嚴重危害人類健康的熱帶傳染病,在全世界人群中具有很高的發(fā)病率和致死率。近年來瘧原蟲多藥抗性株的出現(xiàn)與迅速擴散,以及蚊媒對殺蟲劑耐受性的增加,目前又無有效的抗瘧疫苗,給瘧疾防治工作帶來很大的困難。傳統(tǒng)方法已難于控制該病的流行,迫切需要為瘧疾的防治工作提供新的方法和手段。瘧疾是由蚊蟲叮咬而傳播,瘧原蟲子孢子一旦進入血循環(huán),數(shù)分鐘后便隨血流跨過肝血竇枯否氏細胞(Kuppfer cell,KC)后主動粘附、迅速侵入肝細胞,并進行紅外期增殖,然后進入紅細胞,導(dǎo)致瘧疾的發(fā)作。子孢子侵入肝細胞是瘧疾感染的關(guān)鍵,而阻斷蟲體的侵入,即可防止瘧疾感染,紅外期阻斷疫苗旨在阻斷子孢子侵入宿主肝細胞,防止瘧原蟲紅內(nèi)期的發(fā)育,是瘧疾疫苗的重要組成部分。 唾液腺子孢子感染性受發(fā)育調(diào)節(jié)。正常情況下,中腸內(nèi)卵囊成熟后破裂,子孢子逸出,經(jīng)血淋巴移行到唾液腺后2d,發(fā)育成為具有感染性的子孢子。因此研究瘧原蟲具有侵入肝細胞能力的唾液腺子孢子與不具侵入肝細胞能力的卵囊子孢子之間的差異分子具有重要意義。通過對瘧原蟲子孢子不同時相點侵入相關(guān)基因和蛋白的研究與鑒定,將有助于認識瘧原蟲生長發(fā)育的分子機制,可為瘧疾的防治提供分子理論依據(jù),提供藥物和疫苗新的靶目標(biāo),為采取防治策略戰(zhàn)勝瘧疾打下了堅實的基礎(chǔ)。 在后基因組時代,對基因功能的分析已成為當(dāng)前的主要任務(wù)。而功能基因組學(xué)最大的技術(shù)挑戰(zhàn)是關(guān)于蛋白表達的分析(蛋白質(zhì)組學(xué))。在本實驗室前期的研究中,應(yīng)用蛋白質(zhì)組學(xué)的二維凝膠電泳(two-dimensional gel electrophoresis, 2-DE)技術(shù)、基質(zhì)輔助激光解析電離飛行時間質(zhì)譜(matrix-assisted laser desorption/inoization time of flying mass spectrometry, MALDI- TOF-MS)分析技術(shù)和蛋白質(zhì)信息學(xué)等方法對約氏瘧原蟲卵囊子孢子和唾液腺子孢子這兩個不同發(fā)育時期蟲體的可溶性蛋白進行了分析研究。結(jié)果顯示相對于卵囊子孢子,在唾液腺子孢子中確定比較明顯的差異蛋白點有六個,分別為PyDp1~6(Plasmodium yoelii differential protein 1~6)。這些差異蛋白點主要為一些膜蛋白、表面蛋白和運動相關(guān)的蛋白。推測這些蛋白可能與瘧原蟲子孢子黏附、侵入肝細胞相關(guān),可能為新的阻斷策略和瘧疾多價亞單位疫苗的靶目標(biāo)。因此,加強這些差異表達蛋白的
[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
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R383
本文編號：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
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R383
【參考文獻】
相關(guān)期刊論文 前2條
1 許穎,張錫林,段建華,黃復(fù)生;改進的一維二維電泳法分析瘧原蟲子孢子蛋白質(zhì)的差異表達[J];第三軍醫(yī)大學(xué)學(xué)報;2005年05期
2 許穎,張錫林,段建華,黃復(fù)生;約氏瘧原蟲卵囊、唾液腺子孢子差異表達蛋白的二維電泳-質(zhì)譜分析[J];中國寄生蟲病防治雜志;2005年03期
，本文編號：2221933
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