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

所要求的仪器，一个纳米滴分光光度计和一个梯度PCR机，将一起用于DNA， RNA和蛋白质的分析，这些通常是昂贵的制备和通常可用于非常小体积的实验。这些设备的用途包括用于克隆的cdna的生成和定量以及用于RNA干扰的dsRNA的合成，这是常规的，但对申请人和他们的学员来说是必不可少的。获得资助的项目包括甲壳类动物胚胎滞育、生理休眠状态和增强的抗逆性过程中的基因表达和蛋白质合成分析。分子伴侣，一种在细胞暴露于压力（如热）时保护其他蛋白质的蛋白质，将被检查。翻译后添加的泛素分子对植物细胞蛋白的活性、丰度、运输和定位的影响将被研究。通过添加泛素修饰蛋白质可以调节植物的生长发育、抵御病原菌攻击和抵抗非生物胁迫。因此，该仪器将用于研究细胞、分子和发育生物学的基本方面。这项研究的应用是多方面的。对滞育现象的研究将加强对青蒿作为水产养殖饲料的利用，并有助于消灭能够经历滞育的害虫。分子伴侣Hsp70如何增强水生动物（其中许多用于水产养殖）对细菌性疾病的抵抗力，以及泛素在植物疾病中的作用，将被研究。通过使用所要求的设备，有可能取得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研究问题的实验，这是NSERC支持研究的基石。资助这项适度的建议将使申请人在有效使用拨款资金和进行现代生物学实验方面处于明显的优势。

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.