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

佛罗里达新学院自然科学分部将获得一台实时聚合酶链式反应(PCR)仪器，用于跨学科生物学和分子生物学研究和教学。该仪器可以放大DNA，并使用荧光标记的探针来实时定量DNA和RNA的数量。此外，这项技术需要非常少量的起始材料，这使它比其他测量基因表达的技术具有显著的优势。三名生物和生化教员计划使用这台仪器研究以下三类生物的基因表达：线虫、玉米和珊瑚。在第一个项目中，将利用基因表达监测来研究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，蛋白质RNA解旋酶A(RHA-1)是线虫生殖系染色体外基因阵列沉默和种系正常发育所必需的。实时荧光聚合酶链式反应仪器将被用来研究带有和不带有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生殖系发育所必需的。其他生物的生殖系统似乎也需要基因沉默，因此这个项目可能会对该领域产生广泛影响。在第二个项目中，将研究与重新定位玉米的极性建立有关的基因。本实验室以前的工作指出，在这一过程中，与细胞质乌头酸酶/IRP-1同源物对应的基因上调，以及丝裂原活化蛋白激酶(MAP K)的激活。这些参与者的作用将通过研究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来进一步测试。这样的研究应该有助于了解植物中的IRP-1类似同源物和总体上的重新定位。这项研究可以用来改良玉米作物，以便植株对重新定位(也就是倒伏)做出更快的反应，并减少作物损失。第三个项目涉及健康和患病珊瑚及其共生的虫黄藻的基因表达研究。珊瑚疾病在世界各地都在增加，人们对珊瑚免疫系统或引起珊瑚疾病的生物知之甚少。将分离在感染白色综合症的珊瑚群体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的化验方法。这个项目很有价值，因为珊瑚礁的健康对维持海洋中的生物多样性很重要，而且它应该有助于加深对珊瑚免疫系统的了解。所有这些研究项目都将与从事本科生论文研究和独立研究项目的学生合作进行。该仪器还将用于细胞生物学、遗传学和生物化学的教学实验室课程，开发的课堂项目将传播到更大的社区。购买该仪器将使许多本科生接触到实时PCR技术，这对那些计划攻读科学高级学位的人来说应该是有价值的。

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.