Incubator shaker and accessories

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The instrumentation will be used primarily for the controlled culture of bacteria, generally Escherichia coli. This is an essential procedure for the researcher submitting the application and for others working in the Department of Biology who will have access to the incubator shaker and accessories. Cultured bacteria will be used in cloning cDNAs encoding proteins up-regulated during diapause, and for expression of wild type and mutated eukaryotic proteins which are then purified. These proteins are used in several in vitro assays to investigate the properties of proteins such as molecular chaperones and as antigens for production of antibodies used in protein quantification and localization. The incubator shaker will be used to grow Artemia under controlled temperature and aeration. Gene expression and protein synthesis during Artemia embryo development and diapause, a state of physiological dormancy and enhanced stress tolerance, will be studied. Protein structure/function relationships and the contribution of molecular chaperones to cell survival during diapause are important components of this work. The research employing the incubator shaker will make fundamental contributions to cell, molecular and developmental biology, and there are many applications of the work. Examination of diapause may reveal how insects resist stress, this of importance in agriculture and forestry where the impact of these organisms as pests is exacerbated by the ability to enter diapause and survive harsh environmental conditions. A greater understanding of diapause will enhance the use of Artemia as feed in aquaculture. Dr. MacRae studies the role of HSP27, a small heat shock protein, in breast cancer cell metastasis. Clearly there is potential to make discoveries which have direct consequences to commerce and health through use of the incubator shaker and accessories sought herein. The equipment will also facilitate experiemnts addressing basic questions which are the cornerstone of research programs supported by NSERC. Funding this proposal will put the applicant at a definite advantage in the effective use of grant funds and the ability to conduct modern experimentation in several biological disciplines.