昆蟲的免疫反應(yīng)起始于昆蟲對(duì)不同入侵病原體的精準(zhǔn)識(shí)別。昆蟲通過特定的模式識(shí)別受體(PRRs)與病原體表面的病原體相關(guān)分子模式(PAMPs)相互作用來對(duì)不同病原物進(jìn)行識(shí)別,以區(qū)分自我和非自我,并迅速介導(dǎo)下游的免疫反應(yīng)。C型凝集素(C-typelectin,CTL)是凝集素超家族的重要成員,其作為模式識(shí)別受體(PRRs)參與了昆蟲的免疫防御反應(yīng),在病原物的識(shí)別中具有關(guān)鍵作用。紅棕象甲R(shí)hynchophorus ferrugineus(Olivier)(Coleoptera:Curculionidae)是嚴(yán)重危害棕櫚植物的重要害蟲,并對(duì)棕櫚科植物產(chǎn)業(yè)造成了巨大的經(jīng)濟(jì)損失。隨著對(duì)紅棕象甲免疫機(jī)制和宿主防御的深入研究,CTL在激活免疫反應(yīng)中所起的作用越來越受到人們的關(guān)注�；诖�,本研究克隆分析了一種C型凝集素(RfCTL),其ORF為226 bp,共編碼170個(gè)氨基酸,具有一個(gè)糖識(shí)別區(qū)域(carbohydrate recognition domain,CRD).轉(zhuǎn)錄表達(dá)分析結(jié)果表明,RfCTL在血淋巴和脂肪體中表達(dá)量較高,在注射金黃色葡萄球菌Staphylococcus aureus和大腸桿菌Es...

【文章頁數(shù)】：75 頁

【學(xué)位級(jí)別】：碩士

【文章目錄】：
摘要
ABSTRACT
1. General Introduction
    1.1 CTLs in insect's innate immunity
        1.1.1 CTLs domain organization and structure
        1.1.2 Phylogenetic analysis of CTLs from several insects
        1.1.3 C-type lectins in various Insects
    1.2 Innate immunity in insects
        1.2.1 Cellular immune responses in insects
        1.2.2 Humoral response in insect
    1.3 Pattern recognition receptors (PRRs)
        1.3.1 Peptidoglycan recognition binding proteins (PGRPs)
        1.3.2 Gram-negative binding proteins (GNBPs)
        1.3.3 Hemolin
        1.3.4 Integrin's
    1.4 Signaling pathways in innate immunity
        1.4.1 Toll Pathway in Drosophila melanogaster
        1.4.2 The immune deficiency (IMD) pathway in Drosophila melanogaster
        1.4.3 JAK/STAT pathway
        1.4.4 RNA interference pathway
    1.5 Purpose and significance of this study
2. Materials and Methods
    2.1 Insect collection and rearing
    2.2 RNA extraction, gel electrophoresis, and cDNA synthesis
    2.3 Molecular characterization of isolated template
        2.3.1 Cloning of the target fragment
    2.4 Rapid Amplification of 3 'End Sequence of RfCTL (3' RACE)
        2.4.1 Synthesis of 3 'RACE cDNA
        2.4.2 3'RACE sequence amplification
        2.4.3 Recovery of PCR product and cloning
    2.5 Rapid Amplification of 5 'End Sequence of RfCTL (5'RACE)
        2.5.1 5'RACE sequence amplification
    2.6 Molecular characterization and sequence analysis of RfCTL
    2.7 Phylogenetic analysis
    2.8 Quantitative real-time PCR(qRT-PCR)analysis of the expression profiles of RfCTL indifferent tissues
    2.9 Evaluation of RfCTL in relation to the bacterial challenge
    2.10 RNA interference
        2.10.1 Primer design with T7 promoter
        2.10.2 Preparation of dsRNA template
        2.10.3 Preparation of dsRNA
        2.10.4 dsRNA microinjection
3. Results
    3.1 Characterization and features of RfCTL
    3.2 Phylogenetic analysis of RfCTL with various known CTLs
    3.3 Expression profile of RfCTL across different tissues
    3.4 RfCTL expression is induced by bacterial challenge
    3.5 Verification of RNAi effects on the RfCTL
4. Discussion
Conclusion
References
Acknowledgement



本文編號(hào)：3861379