一种红棕象甲C型凝集素基因的克

发布时间：2023-11-07 20:10

　　昆虫的免疫反应起始于昆虫对不同入侵病原体的精准识别。昆虫通过特定的模式识别受体(PRRs)与病原体表面的病原体相关分子模式(PAMPs)相互作用来对不同病原物进行识别,以区分自我和非自我,并迅速介导下游的免疫反应。C型凝集素(C-typelectin,CTL)是凝集素超家族的重要成员,其作为模式识别受体(PRRs)参与了昆虫的免疫防御反应,在病原物的识别中具有关键作用。红棕象甲Rhynchophorus ferrugineus(Olivier)(Coleoptera:Curculionidae)是严重危害棕榈植物的重要害虫,并对棕榈科植物产业造成了巨大的经济损失。随着对红棕象甲免疫机制和宿主防御的深入研究,CTL在激活免疫反应中所起的作用越来越受到人们的关注。基于此,本研究克隆分析了一种C型凝集素(RfCTL),其ORF为226 bp,共编码170个氨基酸,具有一个糖识别区域(carbohydrate recognition domain,CRD).转录表达分析结果表明,RfCTL在血淋巴和脂肪体中表达量较高,在注射金黄色葡萄球菌Staphylococcus aureus和大肠杆菌Es...

【文章页数】：75 页

【学位级别】：硕士

【文章目录】：
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



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