Instrumentation for Small Volume Applications in DNA, RNA and Protein Analysis

The instrumentation requested, a Nanodrop spectrophotometer and a gradient PCR machine, will be employed together for the analysis of DNA, RNA and proteins which are generally expensive to prepare and often available for experimentation in very small volumes. Uses for the equipment, which encompass the generation and quantitation of cDNAs for cloning and the synthesis of dsRNA for RNA interference, are routine, but essential to the applicants and their trainees. Projects benefiting from the funding include analysis of gene expression and protein synthesis during crustacean embryo diapause, a state of physiological dormancy and enhanced stress tolerance. Molecular chaperones, proteins that protect other proteins during exposure of cells to stress such as heat, will be examined. Effects on activity, abundance, trafficking and localization of plant cell proteins by post-translationally added ubiquitin molecules will be examined. Protein modification by addition of ubiquitins mediates plant growth and development, defense against pathogen attack and resistance to abiotic stress. The instrumentation will therefore be used to study fundamental aspects of cell, molecular and developmental biology. Applications of the research are many. Examination of diapause will enhance the use of Artemia as feed in aquaculture and aid in the eradication of insect pests able to undergo diapause. How the molecular chaperone Hsp70 augments resistance to bacterial diseases in aquatic animals, many of which are used in aquaculture, will be studied as will the role of ubiquitins in plant disease. Through the use of the requested equipment there is potential to make discoveries with direct consequences for commerce and health. The equipment will also facilitate experiments addressing basic research questions, the cornerstone of NSERC supported research. Funding this modest proposal will put the applicants at a definite advantage in the effective use of grant funds and the ability to conduct modern biological experiments.