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首页> 外文学位 >Functional genomics in insect immunology: Identification and characterization of Rhodnius prolixus immune genes.
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Functional genomics in insect immunology: Identification and characterization of Rhodnius prolixus immune genes.

机译：昆虫免疫学中的功能基因组学：Rhodnius prolixus免疫基因的鉴定和表征。

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摘要

Rhodnius prolixus is a bloodfeeding hemipteran insect and a vector of Trypanosoma cruzi, the etiologic agent of Chagas disease. This disease affects over 18 million people in Latin America, and there are no vaccines or efficacious drugs. Because of the unique lifecycle of T. cruzi within the vector, and its avoidance of the hemocoel where the primary immune response occur, we investigated the role of the innate immune response of R. prolixus to pathogens and parasites, including T. cruzi. Insect immunity is based on three principles: pathogen recognition, induction of appropriate regulatory pathways; and the production of molecules to eliminate the pathogens. In this dissertation, I use functional genomics (Suppressive Subtractive Hybridization) to identify and describe genes involved in the immune response of R. prolixus and the implications for T. cruzi. Initially we identified expressed sequence tags (ESTs) corresponding to genes that are differentially expressed in response to parasites and bacteria, and assigned putative gene functions based on sequence similarities. This thesis focuses on four genes of interest including a transcription factor (Dorsal), which may function in the regulation of expression of immune peptides, and three antimicrobial peptides (Defensin, Lysozyme and Prolixin) that serve directly to eliminate pathogens. Transcriptional factor binding motifs (NF-kappaB) present in the promoters of two of these genes (lysozyme and defensin) suggest they are transcriptionally regulated by Dorsal, whereas prolixin is not. We evaluated the temporal and spatial expression profiles of the antimicrobial peptide genes, using real time quantitative PCR, to establish molecular relationships between parasite and vector. Subsequently, we expressed dorsal and prolixin in bacteria and tested their functions. We concluded that invasion of the hemocoel of R. prolixus activates components of the immune system and the production of compounds lethal to T. cruzi, but the pathogen survives by living exclusively in the intestine avoiding vector responses. This study contributes to our overall knowledge of insect immunity, the arsenal of immune molecules available to different insects, and identifies novel and highly conserved immune molecules found in higher and lower insects.;Keywords. Rhodnius prolixus, functional genomics, innate immunity, Lysozyme, Defensin, Dorsal, NF-kappaB.
机译：Rhodnius prolixus是一种吸血的半足类昆虫，是南美锥虫病的病原体克氏锥虫的载体。该疾病在拉丁美洲影响了超过1800万人，并且没有疫苗或有效药物。由于载体中特异克鲁维氏菌的独特生命周期，并且避免了主要免疫反应发生的血细胞，因此我们研究了普氏螺旋体对病原体和寄生虫（包括特异克鲁斯氏菌）的先天免疫应答的作用。昆虫的免疫力基于三个原则：病原体识别，诱导适当的调节途径；并产生消除病原体的分子。在本文中，我使用功能基因组学（抑制性消减杂交）来鉴定和描述与螺旋藻的免疫反应有关的基因及其对克鲁斯锥虫的影响。最初，我们确定了对应于响应寄生虫和细菌而差异表达的基因的表达序列标签（EST），并根据序列相似性分配了推定的基因功能。本论文着眼于四个感兴趣的基因，包括可能在调节免疫肽表达中起作用的转录因子（Dorsal）和直接用于消除病原体的三个抗菌肽（防御素，溶菌酶和脯氨酸）。存在于其中两个基因（溶菌酶和防御素）的启动子中的转录因子结合基序（NF-kappaB）表明它们受背侧转录调控，而脯氨酰胺则不受。我们使用实时定量PCR评估了抗菌肽基因的时间和空间表达谱，以建立寄生虫和载体之间的分子关系。随后，我们在细菌中表达了背和prolixin，并测试了它们的功能。我们得出的结论是，入侵螺旋藻的血细胞激活了免疫系统的组成部分，并产生了对克氏锥虫致死的化合物，但病原体仅通过在肠道中生存而避免了载体反应。这项研究有助于我们全面了解昆虫免疫力，了解不同昆虫可获得的免疫分子库，并鉴定出在高等和低等昆虫中发现的新颖且高度保守的免疫分子。 Rhodnius prolixus，功能基因组学，先天免疫，溶菌酶，防御素，背侧，NF-κB。

著录项

  • 作者

    Ursic Bedoya, Raul.;

  • 作者单位

    Simon Fraser University (Canada).;

  • 授予单位 Simon Fraser University (Canada).;
  • 学科 Genetics.;Entomology.
  • 学位 Ph.D.
  • 年度 2008
  • 页码 211 p.
  • 总页数 211
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 能源与动力工程;
  • 关键词

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