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: 9350368
• 负责人: Robert L Unckless
• 金额: $ 24.58万
• 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/9350368
• 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; Genome; Genomics; Grant; Growth; Health; Human; Immune; Immune response; Infection; Infectious Agent; Innate Immune System; 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; experimental study; fighting; fly; fungus; genetic analysis; genetic manipulation; genome editing; genomic RNA; insight; killings; knockout gene; model development; novel; oral infection; paralogous gene; pathogen; pressure; promoter; public health relevance; 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（由应用程序提供）：抗菌肽作为基因复制抗菌肽（AMPS）进化的模型是真核生物的体液免疫响应的重要组成部分。长期以来，通过重复的基因家族扩展一直被认为是产生进化新颖性的一种手段，尽管从基因重复到新功能的进化过程尚未得到很好的确定。进化的一个罢工特征 AMP是AMP基因家族中基因复制的高率。拟议的研究将在AMP基因家族中使用重复，以更好地了解AMP功能和基因重复的演变。使用AMP研究基因重复可能适用于人类健康，并提供果蝇家庭中基因重复的内置复制。此外，在果蝇中执行高吞吐量实验和遗传操作的能力以及AMP功能中涉及的一组预期表型的存在使系统可拖动。 AIM 1是对物种内部和物种之间的AMP进化的全面分析，以解决选择在基因复制后选择在进化中的作用，以及基因表达与物种和旁系同源物之间的分歧。该分析的一部分将在非常细微的尺度上检查这些AMP的表达，并在空间上（跨组织）和暂时检查。 AIM 1的训练成分包括最先进的人群遗传分析以及精细的基因表达研究。 AIM 2由解决特定AMP旁系同产物的功能，响应于口腔感染和全身感染，使用工程基因敲除并采用多种病原体，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放大器多个副本以测试基因剂量的副本的转基因线，在旁系同源物之间执行启动子交换实验，以测试调节序列的变化如何影响表达在下功能化的途径。 AIM 2中的培训包括使用开发基因组编辑技术来解决这些问题的转基因线的构建。 AIM 3将专门集中于自然种群中隔离的放大器重复，以解决旁系同源的功能的性质。 事件。我们将采用与目标1和2中相同的基本框架检查可变性，基因表达和旁系同源物函数的模式，但专注于这些分离的串联重复。我们还将创建转基因串联重复项，以执行功能性ASSAS控制以实现背景效果。由于AMP基因在基因家族中的重复性的重复性质，我们希望得出有关基因重复进化的结论。拟议的研究的培训部分将完成申请人以前的经验，并将他定位为富有成效的研究职业。康奈尔大学和拉扎罗和克拉克实验室共同拥有资源和专业知识，以确保赠款的培训阶段成功完成。