Refrigerated Centrifuge for Small Volume Applications

本申请中要求的仪器将用于在控制温度下对含有DNA， RNA和蛋白质的小样本（1.5 ml或更少）进行离心。这是申请人、他的学生和其他在生物系工作的人可以使用离心机的常规但必要的程序。本研究将在甲壳类动物阿耳特明（Artemia franciscana）的胚胎中研究滞育（一种生理休眠状态和增强的抗逆性）期间的基因表达和蛋白质合成。从青蒿中提取的DNA、RNA和蛋白质可用于多种功能分析。蛋白质结构/功能关系和分子伴侣在休眠期间保护和折叠其他蛋白质对细胞存活的贡献是这项工作的重要组成部分。因此，冷冻离心机将被用于对细胞、分子和发育生物学做出根本性的贡献。此外，该研究还有许多应用。检查滞育可以揭示昆虫如何抵抗压力，这在农业和林业中很重要，因为这些生物作为害虫的影响由于它们进入滞育和在恶劣环境条件下生存的能力而加剧。对滞育的深入了解将有助于提高青蒿作为水产养殖饲料的使用。应激诱导的Artemia幼虫中分子伴侣HSP70的增强增强了它们对细菌感染的抵抗力，这一观察结果对开发控制重要商业水生无脊椎动物疾病的新程序具有潜在的重要意义。研究热休克小蛋白HSP27在乳腺癌细胞转移中的作用。显然，通过使用冷冻微型离心机，有可能做出对商业和健康有直接影响的发现。该设备还将促进解决基础问题的实验，这些问题是NSERC支持的研究项目的基石。资助这一适度的提案将使申请人在有效使用拨款资金和在几个生物学学科进行现代实验的能力方面处于明确的优势。

The instrument requested in this application will be used for the centrifugation, under controlled temperature, of small samples (1.5 ml or less) containing DNA, RNA and protein. This is a routine but essential procedure for the applicant, his students and others working in the Department of Biology who will have access to the centrifuge. Gene expression and protein synthesis during diapause, a physiological state of dormancy and enhanced stress tolerance, will be studied in embryos of the crustacean, Artemia franciscana. The DNA, RNA and proteins prepared from Artemia are used in several functional assays. Protein structure/function relationships and the contribution of molecular chaperones, which protect and fold other proteins, to cell survival during dormancy are important components of the work. The refrigerated centrifuge will therefore be used to make fundamental contributions to cell, molecular and developmental biology. In addition, there are many applications for the research. 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 their ability to enter diapause and survive harsh environmental conditions. A greater understanding of diapause will enhance the use of Artemia as feed in aquaculture. Stress-induced augmentation of the molecular chaperone HSP70 in Artemia larvae increases their resistance to infection by bacteria, an observation with potential importance in the development of new procedures for the control of disease in commercially important aquatic invertebrate species. The role of HSP27, a small heat shock protein, in breast cancer cell metastasis will be studied. Clearly there is potential to make discoveries which have direct consequences to commerce and health through use of a refrigerated microcentrifuge. The equipment will also facilitate experiments addressing basic questions which are the cornerstone of research programs supported by NSERC. Funding this modest proposal will put the applicant at a definite advantage in the effective use of grant funds and the ability to conduct modern experiments in several biological disciplines.