本文選題：抑制消減雜交 + 大劣按蚊　； 參考：《第三軍醫(yī)大學(xué)》2005年碩士論文

【摘要】：瘧疾是世界上主要的傳染病之一,在全世界人群中具有很高的發(fā)病率和致病率。全球受瘧疾威脅的人口超過(guò)30 億,每年死亡的人數(shù)超過(guò)100 萬(wàn)(WHO:World Malaria Report 2005)。隨著瘧原蟲(chóng)多藥抗性株的出現(xiàn)與迅速擴(kuò)散、蚊媒對(duì)殺蟲(chóng)劑抗性的增加,以及目前尚無(wú)有效的抗瘧疫苗,因此瘧疾的控制工作將面臨巨大的挑戰(zhàn),迫切需要發(fā)展新的瘧疾控制策略。20 世紀(jì)90 年代初,WHO 提出了“遺傳改造蚊媒”的瘧疾防治新策略,而尋找按蚊抗瘧原蟲(chóng)感染相關(guān)基因,克隆、鑒定并分析其功能,是實(shí)現(xiàn)這種新策略的首要工作。 按蚊的先天免疫反應(yīng)主要通過(guò)黑化包被反應(yīng)的激活, NO 和抗菌肽的產(chǎn)生等機(jī)制抑制體內(nèi)瘧原蟲(chóng)的發(fā)育。但是NO 和抗菌肽只能在一定程度上調(diào)節(jié)按蚊體內(nèi)的瘧原蟲(chóng)感染率,而黑化包被反應(yīng)則能完全抑制瘧原蟲(chóng)的感染,是能否阻斷按蚊感染瘧原蟲(chóng)的最主要免疫機(jī)制。當(dāng)瘧原蟲(chóng)表面分子與按蚊的可溶性模式識(shí)別受體結(jié)合而觸發(fā)該反應(yīng),并激活絲氨酸蛋白酶(Serine proteinase,Sp)級(jí)聯(lián),最終導(dǎo)致前酚氧化酶(Prophenoloxidase,PPO)分解激活酚氧化酶(Phenoloxidase,PO),PO 激活后續(xù)成分介導(dǎo)蛋白質(zhì)發(fā)生交聯(lián)并聚合成黑色素從而固定、隔離或殺死入侵的病原體。目前,已從岡比亞按蚊(Anopheles gambiae)克隆到幾種不同的PPO、Sp 和抗菌肽等。但是究竟如何啟動(dòng)了按蚊抗瘧原蟲(chóng)感染的免疫反應(yīng),目前仍不清楚。 大劣按蚊是我國(guó)和東南亞地區(qū)的重要傳瘧蚊媒。研究發(fā)現(xiàn),對(duì)惡性瘧原蟲(chóng)敏感的大劣按蚊可以炓化包被約氏瘧原蟲(chóng)卵囊,所以大劣按蚊-約氏瘧原蟲(chóng)模型是研究按蚊抗瘧原蟲(chóng)感染機(jī)制較好的模型。目前研究按蚊抗瘧原蟲(chóng)感染相關(guān)分子采用的技術(shù)有退化PCR、DD-PCR 等,通過(guò)這些方法獲得基因的數(shù)目有限,而且不能克服基因上調(diào)。本實(shí)驗(yàn)以大劣按蚊-約氏瘧原蟲(chóng)為模型,利用新近建立的SSH 方法構(gòu)建感染約氏瘧原蟲(chóng)大劣按蚊消減文庫(kù),通過(guò)對(duì)差異基因的鑒定和生物信息學(xué)分析,預(yù)測(cè)其在按蚊感染瘧原蟲(chóng)的過(guò)程中所起的作用,并對(duì)與先天免疫相關(guān)的差異基因進(jìn)行半定量實(shí)驗(yàn),分析約氏瘧原蟲(chóng)感染前后其轉(zhuǎn)錄的變化,探討它與按蚊抗瘧原蟲(chóng)感染的先天免疫反應(yīng)的關(guān)系。實(shí)驗(yàn)內(nèi)容和結(jié)果主要包括以下方面:
[Abstract]:Malaria is one of the major infectious diseases in the world. More than 3 billion people worldwide are threatened by malaria, and more than 1 million people die each year at World Malaria Report 2005. With the emergence and rapid spread of multidrug resistant strains of Plasmodium, the increased resistance of mosquito vectors to insecticides, and the absence of effective antimalarial vaccines, malaria control will face enormous challenges. There is an urgent need to develop a new malaria control strategy. In the early 1990s, WHO put forward a new strategy for malaria control with "genetic transformation of mosquito vectors", and searched for genes related to the resistance of Anopheles to Plasmodium infection, cloned, identified and analyzed its function. Is the most important task to realize this new strategy. The innate immune response of Anopheles japonicus is mainly through the activation of melanized envelope reaction, the production of no and antimicrobial peptides, and so on, which inhibits the development of Plasmodium in vivo. However, no and antimicrobial peptides can only regulate the infection rate of malaria parasite in Anopheles mosquitoes to a certain extent, while the melanized coating reaction can completely inhibit the infection of malaria parasites, which is the main immune mechanism to block the infection of Anopheles mosquitoes. When the plasmodium surface molecules bind to the Anopheles soluble pattern recognition receptor, the reaction is triggered and the serine proteinase spa cascade is activated. In the end, Prophenoloxidase (PPO) decomposition activates the subsequent components of Phenoloxidase (PPOP) and mediates the cross-linking of proteins and polymerization into melanin to fix, isolate or kill the invading pathogens. At present, several different PPOSp and antimicrobial peptides have been cloned from Anopheles gambiaeae, Anopheles gambiaeae. But how the Anopheles infestation immune response against Plasmodium infection remains unclear. Anopheles dirus is an important mosquito vector in China and Southeast Asia. It was found that the Anopheles inferioris sensitive to Plasmodium falciparum could be encapsulated with Plasmodium yoelii oocysts, so the Anophele-Plasmodium malarial model was a better model to study the mechanism of Anopheles infestation against Plasmodium falciparum infection. At present, the techniques used to study Anopheles repens against Plasmodium infection include degenerated PCRN DD-PCR. The number of genes obtained by these methods is limited, and the gene upregulation cannot be overcome. Using Anophele-Plasmodium yoelii as a model, a subtractive library of Anopheles dirus infected with Plasmodium yoelii was constructed by using the newly established SSH method. The differential genes were identified and bioinformatics were analyzed. To predict the role of Anopheles in the infection of Plasmodium yoelii, and to analyze the transcription changes of Plasmodium yoelii before and after infection by semi-quantitative experiments of differentially related genes related to innate immunity. To investigate the relationship between Anopheles and the innate immune response of Anopheles to Plasmodium infection. The contents and results of the experiment mainly include the following aspects:
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R383

【共引文獻(xiàn)】

相關(guān)期刊論文 前9條

1 王英;黃復(fù)生;張錫林;徐文岳;段建華;;大劣按蚊絲氨酸蛋白酶AdSP3的組織定位和定量研究[J];成都醫(yī)學(xué)院學(xué)報(bào);2011年02期

2 黃復(fù)生;瘧原蟲(chóng)和蚊相互關(guān)系研究進(jìn)展[J];第三軍醫(yī)大學(xué)學(xué)報(bào);2004年06期

3 徐文岳,黃復(fù)生,夏麗莎,段建華;約氏瘧原蟲(chóng)感染誘導(dǎo)大劣按蚊AdSP1和AdSP3的轉(zhuǎn)錄[J];第三軍醫(yī)大學(xué)學(xué)報(bào);2004年06期

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7 王英;黃復(fù)生;段建華;周桃莉;陳繼德;;大劣按蚊血淋巴中瘧原蟲(chóng)感染相關(guān)蛋白的cDNA克隆[J];寄生蟲(chóng)與醫(yī)學(xué)昆蟲(chóng)學(xué)報(bào);2007年03期

8 張禮生,張青文,蔡青年,徐靜,周明rB;中國(guó)昆蟲(chóng)染色體研究現(xiàn)狀與展望[J];昆蟲(chóng)學(xué)報(bào);2003年06期

9 馬雅軍,瞿逢伊;我國(guó)赫坎按蚊復(fù)合體成員種的rDNA-ITS2區(qū)序列差異及系統(tǒng)發(fā)育分析[J];昆蟲(chóng)知識(shí);2002年03期

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本文編號(hào)：1791482