RUI: Acquisition of DNA Sequencer, Synthesizer and Spectrophotometer

Rochester Institute Of Tech, Rochester NY

Investigators

Abstract

Abstract A grant has been awarded to Dr. Lawlor at Rochester Institute of Technology to purchase three pieces of research equipment: a Genetic Analyzer, a DNA synthesizer and a diode array spectrophotometer. These complementary pieces would form an equipment suite that would facilitate the research programs of four investigators in the departments of biological sciences and chemistry. The research areas are diverse; they range from molecular characterization of immune response molecules in the great apes to creation of bacterial-resistant transgenic plants. They also include the functional characterization of bacterial enzyme systems, as well as the definition of closely related Central-American iguana species. The immune response molecules regulate the activity of natural killer cells, a lymphoid cell population instrumental in detecting virus-infected cells. The transgenic plant project explores the possibility of creating plants with increased production of a naturally occurring molecule that protects against bacterial parasites. The function of threonine dehydrogenase, an essential enzyme in a key metabolic pathway for many bacterial species, will be better understood after molecular characterization of its variant forms. The iguana project is intent on enumerating the species central Mexico and determining how similar they are to one another. The projects, although disparate, share the feature that they are basic biological research projects that would benefit by being able to characterize and sequence the relevant genes. The suite of equipment will permit the requisite DNA sequencing experiments so that the apes can be tested for the presence of the natural killer genes. If present, precise comparisons can be made against those found in human. The plant transgene, created using recombinant DNA techniques, needs to be sequenced to insure proper construction of the gene. Without this quality control assessment, subsequent experiments cannot be undertaken. Preliminary sequence information obtained for gram-negative bacteria, other than E. coli, reveals major differences in the nucleotide sequence for threonine dehydrogenase. These differences will be completely defined by sequencing the gene from the various bacterial species. Earlier field trips to Mexico provided the samples required for DNA sequence analysis of genes, mitochondrial and nuclear, that will form the database for estimating relatedness amongst the species. Although the DNA sequencer is the central equipment item, the DNA synthesizer will allow oligonucleotide production for the sequencing experiments and the spectrophotometer is necessary for accurate estimations of reagent amounts. The scientific contributions from the four projects will be enhanced due to the expanded capability provided by the equipment. Experiments that couldn't be undertaken previously can now be performed in the respective laboratories. It is expected that a better definition of the evolution of the natural killer genes in humans and apes will be one result. Also, testing the hypothesis that transgenic plants are superior to non-transgenic plants in regards to bacterial invasion will be possible. The characterization of the threonine dehydrogenase gene will provide insights as to the regions of the molecule critical for its function. Finally, an understanding of the biogeography of mesoamerica will be increased from the description of the iguana species that are indigenous to that area.