Aquisition of a DNA Sequencer for Plant Molecular Biology Research

点击翻译按钮获取中文摘要

Underpinning much of the current research in plant molecular biology is the ability to routinely sequence genes of biological and economic interest. These genes are being cloned as genomic and cDNA fragment from higher plant species, as well as from the many microbes that act as plant symbionts and pathogens. The ability to isolate Arabidopsis mutants and sequence the genes underlying specific altered phenotypes has introduced a powerful new means of deciphering the molecular mechanisms by which plants grow and develop. Unfortunately, much of the actual research time spent by graduate students and postdoctoral associates in plant molecular biology laboratories is occupied with the rote, laborious procedure by which DNA sequencing `ladders' are electrophoretically separated, detected and analyzed. We estimate that at least 30% of their time is occupied with this procedure, which takes time away from the more critical and intellecutually demanding design of biological experiments. The purpose of this proposal is to obtain federal matching funds to purchase an automated DNA sequencer. This machine will be supervised by Prof. Sussman and operated in his laboratory by a full-time technician with salary provided by state of Wisconsin funds. These automated, fluorescent- based sequencers are widely used in genome sequencing projects, and have been shown to be a cost-effective means of saving time and labor. In addition to time savings, other advantages of the automated technique include the absence of radioactive nuclides and a decreased financial burden associated with their safety and disposal. Major users of this equipment include 8 faculty with federally funded basic research program s in the area of plant molecular biology. The laboratories of these faculty are in close physical proximity but are administered through 5 different academic departments (Departments of Horticulture, Agronomy, Biochemistry and Plant Pathology in the College of Agricultural and Life Sciences; Department of Botany in the College of Letters and Science). Minor users include several newly hired `applied' geneticists with active research programs using recombinant DNA techniques for more efficient breeding of improved crop plants.