Antimicrobial peptides as models for the evolution of gene duplication

抗菌肽作为基因复制进化的模型

• 批准号: 9330300
• 负责人: Robert L Unckless
• 金额: $ 24.21万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9330300
• 关键词: Address; Amino Acid Sequence; Animals; Bacteria; Bacterial Infections; Biological; Biological Assay; Biological Models; Characteristics; Code; Communities; Complement; Conflict (Psychology); Copy Number Polymorphism; Data; Development; Dissection; Drosophila genus; Employee Strikes; Engineered Gene; Engineering; Ensure; Eukaryota; Event; Evolution; Family; Gene Conversion; Gene Dosage; Gene Duplication; Gene Expression; Gene Family; Gene Proteins; Genes; Genetic; Grant; Growth; Health; Human; Immune; Immune response; Immune system; Infection; Infectious Agent; Isoptera; Knock-out; Link; Measures; Methods; Modeling; Molecular; Nature; Open Reading Frames; Organ; Organism; Pattern; Phase; Phenotype; Plants; Play; Population; Population Genetics; Positioning Attribute; Process; Proteins; Publishing; RNA Sequences; Research; Resistance to infection; Resolution; Resources; Role; Route; Scientist; Shapes; System; Systemic infection; Techniques; Technology; Testing; Theoretical model; Time; Tissues; Training; Transgenic Organisms; Universities; Variant; Virus; Work; antimicrobial peptide; base; career; career development; combat; dosage; duplicate genes; experience; fighting; fly; functional genomics; fungus; genetic analysis; genetic manipulation; genome editing; genome sequencing; genomic RNA; insight; killings; model development; novel; oral infection; paralogous gene; pathogen; pressure; promoter; protein expression; research study; response; screening; skills; theories; tool

 DESCRIPTION (provided by applicant): Antimicrobial peptides as models for the evolution of gene duplication Antimicrobial peptides (AMPs) are a vital part of the humoral immune response for eukaryotes. Gene family expansion through duplication has long been recognized as a means of generating evolutionary novelty though the evolutionary processes leading from gene duplication to novel function is not well established. One striking characteristic of the evolution of AMPs is the high rate of gene duplication in AMP gene families. The proposed research will use duplications in AMP gene families to better understand both AMP function and the evolution of gene duplication. Using AMPs to study gene duplication may have applicability to human health and provides built-in replication of gene duplications across families in Drosophila. Furthermore, the ability to perform high throughput experiments and genetic manipulations in Drosophila and the existence of a set of expected phenotypes involved in AMP function make the system tractable. Aim 1 is a comprehensive analysis of AMP evolution within and between species to address the role that selection plays in evolution after gene duplication and how gene expression diverges with between species and paralogs. Part of this analysis will examine expression of these AMPs on a very fine scale both spatially (across tissues) and temporally. The training component of Aim 1 includes state-of-the art population genetic analyses as well as fine scale gene expression studies. Aim 2 consists addresses the function of specific AMP paralogs in response to both oral infection and systemic infection using engineered gene knockouts and employing a large variety of pathogens since we have evidence that AMP immune response is pathogen-specific. Based on the results from this experiment, we will create transgenic lines with multiple copies of a particular AMP to test the effects of gene dosage, perform promoter swapping experiments between paralogs to test how changes in regulatory sequence might influence expression en route to subfunctionalization. Training in Aim 2 consists of the construction of transgenic lines using developing genome editing technologies to address these questions. Aim 3 will focus specifically on AMP duplicates segregating in natural populations to address the nature of paralog function shortly after the initial duplication event. We will employ the same basic framework as in Aims 1 and 2 examining patterns of variability, gene expression and paralog function but focusing on these segregating tandem duplicates. We will additionally create transgenic tandem duplicates to perform functional assays controlling for background effects. Because of the replicated nature of AMP gene duplicates across gene families, we expect to draw conclusions about the evolution of gene duplication. The training portion of the proposed research will complement the applicant's previous experience and position him for a productive research career. Cornell University and the Lazzaro and Clark labs together have the resources and expertise to ensure the successful completion of the training phase of the grant.

 描述（由申请人提供）：作为基因复制进化模型的抗菌肽抗菌肽（AMP）是真核生物体液免疫反应的重要组成部分。尽管从基因复制到新功能的进化过程尚未完全确定，但通过复制进行基因家族扩张长期以来一直被认为是产生进化新颖性的一种手段。进化的一个显着特征 AMP 的主要特点是 AMP 基因家族中基因重复率很高。拟议的研究将利用 AMP 基因家族中的重复来更好地了解 AMP 功能和基因重复的进化。使用 AMP 研究基因重复可能适用于人类健康，并提供果蝇科中基因重复的内置复制。此外，在果蝇中进行高通量实验和遗传操作的能力以及参与 AMP 功能的一组预期表型的存在使得该系统易于处理。目标 1 是对物种内和物种间 AMP 进化的全面分析，以解决基因复制后选择在进化中所起的作用，以及基因表达如何在物种和旁系同源物之间产生差异。该分析的一部分将在空间（跨组织）和时间上以非常精细的尺度检查这些 AMP 的表达。目标 1 的培训部分包括最先进的群体遗传分析以及精细规模的基因表达研究。目标 2 包括使用工程基因敲除和使用多种病原体来解决特定 AMP 旁系同源物对口腔感染和全身感染的功能，因为我们有证据表明 AMP 免疫反应是病原体特异性的。根据该实验的结果，我们将创建具有特定 AMP 的多个拷贝的转基因系，以测试基因剂量的影响，在旁系同源物之间进行启动子交换实验，以测试调控序列的变化如何影响亚功能化过程中的表达。目标 2 的培训包括使用开发的基因组编辑技术构建转基因品系来解决这些问题。目标 3 将特别关注自然群体中 AMP 复制品的分离，以解决初始复制后不久的旁系同源功能的性质 事件。我们将采用与目标 1 和 2 相同的基本框架来检查变异性、基因表达和旁系同源功能的模式，但重点关注这些分离的串联重复。我们还将另外创建转基因串联复制品，以进行控制背景效应的功能测定。由于跨基因家族的 AMP 基因重复的复制性质，我们期望得出有关基因重复进化的结论。拟议研究的培训部分将补充申请人以前的经验，并使他能够从事富有成效的研究生涯。康奈尔大学以及拉扎罗和克拉克实验室共同拥有资源和专业知识，以确保成功完成资助的培训阶段。