【摘要】：媒介昆蟲(chóng)嚴(yán)重危害人類健康,據(jù)估計(jì)傳染病中2/3由媒介昆蟲(chóng)傳播。世界史上許多危害嚴(yán)重的蟲(chóng)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(wàn)人的生命。當(dāng)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(chóng)媒傳染病�；瘜W(xué)防治一直是媒介綜合治理的主要方法。殺蟲(chóng)劑的大量、連續(xù)使用導(dǎo)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(chóng)產(chǎn)生了抗藥性,包括主要的媒介昆蟲(chóng),其中109種(亞種)蚊對(duì)一種或多種殺蟲(chóng)劑產(chǎn)生了抗性。殺蟲(chóng)劑抗性直接影響著蟲(chóng)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(jī)制并進(jìn)行適宜治理實(shí)乃當(dāng)務(wù)之急。本研究擬從克隆抗藥性相關(guān)基因入手,并對(duì)其進(jìn)行功能鑒定,然后運(yùn)用RNAi技術(shù)抑制抗藥性基因,為探討最終治理媒介抗藥性奠定基礎(chǔ)。為此,本文進(jìn)行了以下幾個(gè)方面的研究。 第一部分 淡色庫(kù)蚊溴氰菊酯抗性相關(guān)基因的分子克隆與功能研究 研究一 淡色庫(kù)蚊細(xì)胞色素P450 CYP6F1基因的分子克隆、序列分析及表達(dá)差異鑒定 為克隆淡色庫(kù)蚊細(xì)胞色素P450(CYP6F1)基因并對(duì)其進(jìn)行表達(dá)差異的鑒定,本研究采用RT-PCR技術(shù)和RACE策略,從淡色庫(kù)蚊(Culex pipiens pallens)對(duì)溴氰菊酯抗性品系4齡幼蟲(chóng)中克隆細(xì)胞色素P450基因(CYP6F1),用相應(yīng)的軟件進(jìn)行生物信息學(xué)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(dá)水平及對(duì)蚊各期CYP6F1進(jìn)行表達(dá)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(zhǎng)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(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2005
【分類號(hào)】：R346

【引證文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

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

相關(guān)碩士學(xué)位論文 前4條

1 祝全東;桉樹(shù)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(xué);2011年

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

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

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

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本文編號(hào)：2140758