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

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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.