蚊抗药性相关新基因—mdr的初步研究

发布时间：2018-04-19 20:39

本文选题：淡色库蚊 + 抗药性　；参考：《南京医科大学》2010年硕士论文

【摘要】： 蚊媒病是当今严重危害人类健康的一类常见病,蚊媒的化学防制是控制蚊媒病的一个重要手段。然而随着杀虫剂的长期、大量施用,蚊媒的杀虫剂抗性也随之产生和发展。杀虫剂抗性已成为蚊媒病防治工作中的一大障碍。发现和研究新的抗药性相关基因将有助于阐明抗药性的分子基础和寻找新的治理抗药性的靶标。本实验室在前期实验中发现,多药耐药蛋白在蚊对杀虫剂抗性细胞高表达。在此基础上,本实验通过实时定量PCR法比较淡色库蚊(Culex pipiens pallens)多药耐药基因(multidrug resistance,mdr)在溴氰菊酯抗性品系和敏感品系中的表达差异;采用PCR法扩增淡色库蚊mdr完整开读框,用BlastX软件将该基因推导编码的氨基酸序列与蛋白质公共数据库Swissprot进行分析,用Clustal1.83软件作序列比对,以MEGA 4.0软件进行聚类分析,构建进化树。结果显示,淡色库蚊mdr在抗性品系中高表达,是敏感品系的42.10倍(p0.005);获得的淡色库蚊mdr完整开读框序列为3936bp,编码1311个氨基酸;与致倦库蚊(Cx.quinquefasciatus)、埃及伊蚊(Aedes aegypti)和冈比亚按蚊(Anopheles gambiae)的同源性分别为97%,74%,73%;通过MEGA4.0软件进行聚类分析、构建进化树及序列归一化距离分析,淡色库蚊的mdr同致倦库蚊的距离最近(0.004),其次是埃及伊蚊(0.161)和冈比亚按蚊(0.201); 以上结果提示,mdr可能与蚊抗药性相关,值得进一步研究。本研究结果为进一步探讨蚊抗药性的分子机制和建立抗药性的分子检测方法提供了新的科学依据,具有重要的理论意义和潜在的实用价值。
[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.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R184

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