本文選題：淡色庫(kù)蚊 + 抗藥性�。� 參考：《南京醫(yī)科大學(xué)》2010年碩士論文

【摘要】： 蚊媒病是當(dāng)今嚴(yán)重危害人類(lèi)健康的一類(lèi)常見(jiàn)病,蚊媒的化學(xué)防制是控制蚊媒病的一個(gè)重要手段。然而隨著殺蟲(chóng)劑的長(zhǎng)期、大量施用,蚊媒的殺蟲(chóng)劑抗性也隨之產(chǎn)生和發(fā)展。殺蟲(chóng)劑抗性已成為蚊媒病防治工作中的一大障礙。發(fā)現(xiàn)和研究新的抗藥性相關(guān)基因?qū)⒂兄陉U明抗藥性的分子基礎(chǔ)和尋找新的治理抗藥性的靶標(biāo)。 本實(shí)驗(yàn)室在前期實(shí)驗(yàn)中發(fā)現(xiàn),多藥耐藥蛋白在蚊對(duì)殺蟲(chóng)劑抗性細(xì)胞高表達(dá)。在此基礎(chǔ)上,本實(shí)驗(yàn)通過(guò)實(shí)時(shí)定量PCR法比較淡色庫(kù)蚊(Culex pipiens pallens)多藥耐藥基因(multidrug resistance,mdr)在溴氰菊酯抗性品系和敏感品系中的表達(dá)差異;采用PCR法擴(kuò)增淡色庫(kù)蚊mdr完整開(kāi)讀框,用BlastX軟件將該基因推導(dǎo)編碼的氨基酸序列與蛋白質(zhì)公共數(shù)據(jù)庫(kù)Swissprot進(jìn)行分析,用Clustal1.83軟件作序列比對(duì),以MEGA 4.0軟件進(jìn)行聚類(lèi)分析,構(gòu)建進(jìn)化樹(shù)。 結(jié)果顯示,淡色庫(kù)蚊mdr在抗性品系中高表達(dá),是敏感品系的42.10倍(p0.005);獲得的淡色庫(kù)蚊mdr完整開(kāi)讀框序列為3936bp,編碼1311個(gè)氨基酸;與致倦庫(kù)蚊(Cx.quinquefasciatus)、埃及伊蚊(Aedes aegypti)和岡比亞按蚊(Anopheles gambiae)的同源性分別為97%,74%,73%;通過(guò)MEGA4.0軟件進(jìn)行聚類(lèi)分析、構(gòu)建進(jìn)化樹(shù)及序列歸一化距離分析,淡色庫(kù)蚊的mdr同致倦庫(kù)蚊的距離最近(0.004),其次是埃及伊蚊(0.161)和岡比亞按蚊(0.201); 以上結(jié)果提示,mdr可能與蚊抗藥性相關(guān),值得進(jìn)一步研究。本研究結(jié)果為進(jìn)一步探討蚊抗藥性的分子機(jī)制和建立抗藥性的分子檢測(cè)方法提供了新的科學(xué)依據(jù),具有重要的理論意義和潛在的實(shí)用價(jià)值。
[Abstract]:Mosquito vector disease is a common disease that seriously endangers human health. The chemical control of mosquito vector is an important means to control mosquito vector disease. However, with the long-term application of insecticides, mosquito-borne insecticide resistance also emerged and developed. Insecticide resistance has become a major obstacle in mosquito vector disease control. The discovery and study of new resistance related genes will help to clarify the molecular basis of resistance and to find new targets for the treatment of drug resistance. In our laboratory, we found that MDR protein was highly expressed in insecticide resistant cells. On this basis, the expression difference of multidrug resistance gene of Culex pipiens pallensens was compared by real-time quantitative PCR in deltamethrin resistant and sensitive strains, and the complete open reading frame of Culex pallensens pallensens was amplified by PCR method. The deduced amino acid sequence of the gene was analyzed with the common protein database Swissprot, the sequence was compared with Clustal1.83 software, and the phylogenetic tree was constructed by clustering analysis with MEGA 4.0 software. The results showed that the mdr expression of Culex pipiens pallens p0.005 was 42.10 times higher than that of sensitive strains, and the complete open reading frame sequence of mdr was 3936 BP, encoding 1311 amino acids. The homology with Cx.quinquefasciatusus, Aedes aegyptid and Anopheles gambiae were 9774 and 7333, respectively. The phylogenetic tree and sequence normalization distance analysis were constructed by MEGA4.0 software. The distance between mdr of Culex pipiens pallens and Culex pipiens quinquefasciatus is 0.004, followed by Aedes aegypti 0.161) and Anopheles gambiae 0.201. These results suggest that MDR may be related to mosquito resistance, and it is worthy of further study. The results of this study provide a new scientific basis for further study on the molecular mechanism of mosquito resistance and the establishment of molecular detection methods for drug resistance, which has important theoretical significance and potential practical value.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R184

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