【摘要】：媒介昆蟲嚴重危害人類健康,據(jù)估計傳染病中2/3由媒介昆蟲傳播。世界史上許多危害嚴重的蟲媒傳染病如鼠疫、斑疹傷寒、黃熱病、瘧疾等都曾造成廣泛的流行并奪去成千上萬人的生命。當今世界隨著人類交往的日益頻繁,發(fā)現(xiàn)許多新的和重新出現(xiàn)的蟲媒傳染病。化學防治一直是媒介綜合治理的主要方法。殺蟲劑的大量、連續(xù)使用導致了媒介抗藥性的發(fā)生和發(fā)展。目前,世界范圍內(nèi)至少已有504種昆蟲產(chǎn)生了抗藥性,包括主要的媒介昆蟲,其中109種(亞種)蚊對一種或多種殺蟲劑產(chǎn)生了抗性。殺蟲劑抗性直接影響著蟲媒傳染病的流行和重新流行,因此,研究媒介抗藥性產(chǎn)生的機制并進行適宜治理實乃當務之急。本研究擬從克隆抗藥性相關基因入手,并對其進行功能鑒定,然后運用RNAi技術抑制抗藥性基因,為探討最終治理媒介抗藥性奠定基礎。為此,本文進行了以下幾個方面的研究。 第一部分 淡色庫蚊溴氰菊酯抗性相關基因的分子克隆與功能研究 研究一 淡色庫蚊細胞色素P450 CYP6F1基因的分子克隆、序列分析及表達差異鑒定 為克隆淡色庫蚊細胞色素P450(CYP6F1)基因并對其進行表達差異的鑒定,本研究采用RT-PCR技術和RACE策略,從淡色庫蚊(Culex pipiens pallens)對溴氰菊酯抗性品系4齡幼蟲中克隆細胞色素P450基因(CYP6F1),用相應的軟件進行生物信息學分析,并用定量PCR和半定量RT-PCR分析敏感、抗性品系蚊的CYP6F1表達水平及對蚊各期CYP6F1進行表達差異鑒定。結(jié)果表明,成功克隆出CYP6F1基因及其等位基因CYP6F1v1。CYP6F1基因全長1639 bp,
[Abstract]:Vector insects are a serious hazard to human health, and it is estimated that 2 / 3 of infectious diseases are transmitted by vector insects. In the history of the world, many serious insect-borne infectious diseases such as plague, typhus, yellow fever and malaria have caused widespread epidemic and killed thousands of people. With the increasing frequency of human communication, many new and re-emerging insect-borne infectious diseases have been discovered in the world today. Chemical control has been the main method of comprehensive media management. The extensive and continuous use of insecticides has led to the development and development of vector resistance. At present, at least 504 species of insects have developed resistance to insecticides worldwide, including the major vector insects, 109 of which (subspecies) have developed resistance to one or more insecticides. Insecticide resistance has a direct impact on the prevalence and re-prevalence of insect-borne infectious diseases. Therefore, it is urgent to study the mechanism of vector resistance and to manage it properly. The purpose of this study was to clone and identify the genes related to drug resistance, and then to use RNAi technique to inhibit the resistance genes, and to lay a foundation for the final treatment of drug resistance vector. Therefore, this paper has carried on the following several aspects research. Part I Molecular cloning and functional study of Deltamethrin Resistance related Gene of Culex pipiens pallens; Molecular cloning of cytochrome P450 CYP6F1 gene from Culex pipiens pallens pallens Sequence analysis and differential expression identification were used to clone and express the cytochrome P450 (CYP6F1) gene of Culex pipiens pallens. RT-PCR technique and RACE strategy were used in this study. The cytochrome P450 (CYP6F1) gene was cloned from the (Culex pipiens pallens) of Culex pipiens pallens to the 4th instar larvae of the deltamethrin resistant strain. The bioinformatics analysis was carried out with the corresponding software, and the sensitivity was analyzed by quantitative PCR and semi-quantitative RT-PCR. The expression level of CYP6F1 and the differential expression of CYP6F1 in different stages of mosquito were identified. The results showed that the CYP6F1 gene and its allele CYP6F1v1.CYP6F1 gene were cloned successfully with a total length of 1639 BP.
【學位授予單位】：南京醫(yī)科大學
【學位級別】：博士
【學位授予年份】：2005
【分類號】：R346

【引證文獻】

相關博士學位論文 前1條

1 賀驥;淡色庫蚊核糖體蛋白L22基因(RPL22)克隆及其與溴氰菊酯抗性關系的研究[D];南京醫(yī)科大學;2009年

相關碩士學位論文 前4條

1 祝全東;桉樹CAD、COMT基因的RNAi載體構(gòu)建及煙草轉(zhuǎn)化研究[D];中南林業(yè)科技大學;2011年

2 劉應保;淡色庫蚊氯菊酯抗性相關基因XND-P450全長cDNA克隆及序列分析[D];西北農(nóng)林科技大學;2010年

3 溫雪梅;淡色庫蚊溴氰菊酯抗性基因XN-P450的克隆、分析及CYP6F1的表達[D];西北農(nóng)林科技大學;2012年

4 王曉宇;淡色庫蚊氯菊酯抗性相關actin基因和P450基因全長cDNA克隆及序列分析[D];西北農(nóng)林科技大學;2012年

，

本文編號：2140758