Nematicidal Chalcones: A Genetic Approach to Uncovering the Action Mechanism

标题：杀线虫查尔酮：揭示作用机制的遗传学方法 项目摘要/摘要： 寄生线虫是植物和动物(包括人类)的敌人。这些寄生虫有一种 对医学、兽医和农业产生巨大影响，造成人类感染和死亡 (见世卫组织“土壤传播蠕虫感染”情况说明书)和每年的重大货币损失。例如, 据估计，全世界有1570亿美元的损失(Li等人，2007年)，原因是植物寄生虫的侵扰 线虫。因此，开发新的杀线虫剂并了解其作用机理具有重要意义 最重要的是；线虫控制的解决方案很可能来自于使用鸡尾酒 杀线虫剂，可以通过不同的方式控制线虫。我们实验室之前的工作确定了两个 有机查尔酮(查尔酮17和查尔酮30)对线虫及其植物的有效杀线虫作用 寄生线虫南方根结线虫(Saeed等人)2012年，和Calderón-Urrea等人，未发表的结果)。 此外，这些查尔酮对土壤微生物或人类细胞都没有显著影响。 (Calderón-Urrea等人，未发表的结果)。然而，这些查尔酮的作用机制尚不清楚。 有可能这些查尔酮针对的是基本蛋白质(酶)，而这些蛋白质(酶)是由 查尔康斯。如果是这样的话，改变查尔酮与上述蛋白质的结合可能是一个强有力的工具 目的：阐明查尔酮的作用机制。我们假设“对查尔酮的易感性是 由于查尔酮对蛋白质产物的作用，可能是一种酶，从而导致基因突变 编码该蛋白质的(或基因)将使查尔酮作用无效“。我们正在通过以下方式检验这一假设 对诱变的线虫进行正向遗传筛选，并对分离的线虫进行特征分析 变种人。该项目的具体目标是：1)测试对查尔酮17和30的抗性并鉴定 克服查尔酮效应的突变体；2)查尔酮抗性突变体的特征；以及3)鉴定 查尔酮抗性突变体所涉及的DNA序列。私家侦探将利用他丰富的经验 招募和培训学生，特别是来自科学方面代表性不足的群体的学生，以积极参与 在这个项目的执行过程中。

Title: Nematicidal chalcones: a genetic approach to uncovering the action mechanism Project Summary/Abstract: Parasitic Nematodes (PNs) are enemy to both plant and animals (including humans). These parasites have a tremendous impact on medicine, veterinary medicine, and agriculture, causing human infections and mortality (see WHO “Soil-transmitted helminth infections” factsheet) and significant annual monetary losses. For example, there is an estimated loss of $157 billion worldwide (Li et al., 2007) due to infestation by plant parasitic nematodes. Therefore, developing new nematicidal agents and understanding their mechanism of action is of paramount importance; it is likely that a solution to nematode control will come from the use of cocktails of nematicidal agents, which can control nematodes in different ways. Previous work in our lab identified two organic chalcones (chalcones 17 and 30) as effective nematicidals on Caenorhabditis elegans and the plant parasitic nematode Meloidogyne incognita (Saeed, et al. 2012, and Calderón-Urrea, et al., unpublished results). Furthermore, these chalcones have no significant effects on either soil microorganisms or human cells (Calderón-Urrea, et al., unpublished results). However, the mechanism of action of these chalcones is not known. It is possible that these chalcones target essential proteins (enzymes), which are rendered infective by the chalcones. If this is the case, alterations in the binding of the chalcones to said proteins can be a powerful tool to elucidate the mechanism of action of the chalcones. We hypothesize that “the susceptibility to chalcones is due to the action of the chalcone on a protein product, perhaps an enzyme, and therefore mutations on the gene (or genes) encoding that protein will render the chalcone action ineffective”. We are testing this hypothesis by conducting a forward genetic screen on mutagenized C. elegans nematodes, and characterizing the isolated mutants. The specific objectives of this project are: 1) To test resistance to chalcones 17 and 30 and identification of mutants overcoming chalcone effects; 2) Characterization of chalcone resistant mutants; and 3) Identification of the DNA sequences involved in the chalcone resistant mutants. The PI will use his extensive experience in recruiting and training students, particularly from underrepresented groups in sciences, to actively engage them in the execution of this project.