Acquisition of a Real-Time PCR Machine for Biology Research and Teaching

购置实时荧光定量PCR仪用于生物学研究和教学

• 批准号: 0520607
• 负责人: Katherine Walstrom
• 金额: $ 4.32万
• 依托单位: New College of Florida
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-15 至 2008-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0520607&HistoricalAwards=false
• 关键词: Acquisition; Real; Time; PCR; Machine

The Division of Natural Sciences at New College of Florida will acquire a real-time polymerase chain reaction (PCR) instrument for use in interdisciplinary biology and molecular biology research and teaching. This instrument amplifies DNA and uses fluorescently-labeled probes to allow quantification of the amount of DNA and RNA present in real time. In addition, the technique requires very small amounts of starting material, giving it a significant advantage over other techniques to measure gene expression. Three biology and biochemistry faculty members plan to use this instrument to study gene expression in the following three groups of organisms: C. elegans, maize, and corals. In the first project, monitoring of gene expression will be used to study how development of the reproductive system in the worm C. elegans depends on gene silencing. This lab found previously that the protein RNA helicase A (RHA-1) is required for silencing of extrachromosomal gene arrays in the C. elegans germline and for proper germline development. The real-time PCR instrument will be used to investigate the expression of endogenous genes in worms with and without RHA-1 to determine how gene silencing is required for proper germline development. The reproductive systems of other organisms appear to also require gene silencing, so this project could have a broad impact on the field. In the second project, genes involved in the establishment of polarity in reoriented maize will be studied. Previous work in this lab points to an upregulation of a gene corresponding to a cytoplasmic aconitase/IRP-1 homologue as well to mitogen activated protein kinase (MAP kinase) activation in this process. The roles of these players will be tested further by studying the expression levels of IRP-1 and other genes over a detailed reorientation time course. Such studies should contribute to knowledge of both IRP-1-like homologues in plants and reorientation in general. This study could be used to improve maize crops so that the plants respond more quickly to reorientation (a.k.a., lodging) and reduce crop loss. The third project involves gene expression studies of healthy and diseased corals and their symbiotic zooxanthellae. Coral disease is on the increase throughout the world, and little is known about the coral immune system or about the organisms that cause coral diseases. Coral genes will be isolated that are differentially expressed in coral colonies infected with white syndrome to determine how corals respond to infection. The goal is to develop an assay to assess coral health. This project is valuable because the health of coral reefs is important for maintaining organismic diversity in the ocean, and because it should lead to an increased understanding of the immune systems of corals.All of these research projects will be carried out in collaboration with students working on their undergraduate thesis research and independent study projects. The instrument will also be used in the cell biology, genetics, and biochemistry teaching laboratory courses, and the class projects developed will be disseminated to the larger community. Acquisition of the instrument will expose many undergraduate students to the real time PCR technique, which should be valuable for those who plan to pursue an advanced degree in the sciences.

佛罗里达新学院自然科学系将获得一台实时聚合酶链反应（PCR）仪器，用于跨学科生物学和分子生物学的研究和教学。该仪器扩增DNA，并使用荧光标记探针，允许实时定量DNA和RNA的数量。此外，该技术需要非常少量的起始材料，这使得它比其他测量基因表达的技术具有显著的优势。三名生物学和生物化学教员计划使用该仪器研究以下三组生物的基因表达：秀丽隐杆线虫、玉米和珊瑚。在第一个项目中，基因表达监测将用于研究秀丽隐杆线虫生殖系统的发育如何依赖于基因沉默。本实验室先前发现，RNA解旋酶A （RHA-1）蛋白是秀丽隐杆线虫种系染色体外基因阵列沉默和正常种系发育所必需的。real-time PCR仪器将用于研究携带和不携带RHA-1的蠕虫体内内源基因的表达，以确定基因沉默是如何为正常的种系发育所必需的。其他生物的生殖系统似乎也需要基因沉默，因此该项目可能对该领域产生广泛的影响。在第二个项目中，将研究与重定向玉米极性建立有关的基因。本实验室先前的工作指出，在这一过程中，一个与细胞质乌头酸酶/IRP-1同源物相对应的基因上调，以及有丝分裂原活化蛋白激酶（MAP激酶）的活化。通过研究IRP-1和其他基因在详细的重新定位时间过程中的表达水平，将进一步测试这些参与者的作用。这些研究应该有助于了解植物中irp -1样同源物和一般的定向。该研究可用于改进玉米作物，使植物对重新定向（即倒伏）作出更快的反应，减少作物损失。第三个项目涉及健康和患病珊瑚及其共生虫黄藻的基因表达研究。珊瑚疾病在世界范围内呈上升趋势，而人们对珊瑚的免疫系统或引起珊瑚疾病的生物体知之甚少。将分离在感染白色综合症的珊瑚群落中差异表达的珊瑚基因，以确定珊瑚对感染的反应。其目标是开发一种评估珊瑚健康状况的方法。这个项目很有价值，因为珊瑚礁的健康对维持海洋生物多样性很重要，因为它应该会增加对珊瑚免疫系统的了解。所有这些研究项目将与从事本科论文研究和独立研究项目的学生合作进行。该仪器也将用于细胞生物学、遗传学和生物化学的教学实验课程，并且开发的课堂项目将传播到更大的社区。该仪器的获得将使许多本科生接触到实时PCR技术，这对于那些计划在科学领域攻读高级学位的人来说应该是有价值的。