Crystallization Robot for the Structural Biology Resource Center at Rockefeller

描述（由申请人提供）：本提案申请资金购买机器人，为洛克菲勒大学结构生物学资源中心建立结晶实验。该机器人将帮助研究人员获得涉及细菌转录及其调控、细菌双组分组氨酸激酶信号传导、细菌发病机制、丙型肝炎病毒复制和真核细胞分裂的生物大分子的衍射质量晶体。在x射线结构测定中，获得晶体常常是限制速度的步骤。该机器人将通过减少所需的时间和样品量，大大提高结晶试验的效率。反过来，这将提高美国国立卫生研究院资助的研究的成功率，并将提高美国国立卫生研究院资助的x射线设备已经在资源中心的利用率。由于与手动设置相反，机器人所需的材料数量较少，因此机器人还将允许对项目进行广泛的结晶试验，否则这些试验是不可能的。此外，结晶实验产生的废物少得多，对环境的影响也小得多。要求的结晶机器人是Innovadyne公司的Screenmaker96+8 - 145000美元。这个机器人可以执行三种不同的过程，以建立结晶实验。步骤1：将储层溶液从稀疏基质原液转移到结晶盘中的96个位置；步骤2：将该储层的纳升体积沉积到滴位置，并将蛋白质的纳升体积沉积到同一位置；步骤3：通过改变引线条件中的两个组件来定制网格，从而为改进晶体生长创造新的机会。Screenmaker96+8的另一个非常重要的特点是，放置液滴（沉淀溶液和蛋白质）所需的时间明显少于其他结晶机器人，从而将脱水时间减少至少一半。结晶机器人将被安置在新成立的结构生物学资源中心，该中心由洛克菲勒大学研究支持办公室支持。本中心对机器人的使用进行监督，对使用者进行培训，维护仪器处于最佳工作状态，并对机器人的公平、相关、高效使用负责。所需额外零件：电脑$2450，安装$4000，两个巢式冷水机系统$10450，敏感样品和延长保修期$32000。（这里所报的价格不反映10%的学术折扣适用于预算中要求的总额，请参阅Equipment.doc中的报价）。

DESCRIPTION (provided by applicant): This proposal requests funds to purchase a robot to set up crystallization experiments for the Structural Biology Resource Center at The Rockefeller University. The robot will aid researchers in obtaining diffraction quality crystals of biological macromolecules involved in bacterial transcription and its regulation, bacterial two-component histidine kinase signaling, bacterial pathogenesis, hepatitis C virus replication, and eukaryotic cell division. Obtaining crystals is often the rate-limiting step in X-ray structure determination. The robot will greatly increase the efficiency of crystallization trials by reducing the amount of time and sample required. This, in turn, will increase the success rate of NIH-funded research, and will increase utilization of the NIH-funded X-ray equipment already in the Resource Center. Due to the smaller quantities of materials required for the robot as opposed to manual setup, the robot will also allow extensive crystallization trials for projects that would otherwise not be possible. In addition, crystallization experiments will have a smaller impact on the environment by generating much less waste. The crystallization robot requested is the Screenmaker96+8 from Innovadyne - $145000. This robot can perform three distinct processes needed to set up crystallization experiments. Step 1: transfer of reservoir solutions from the sparse matrix stocks to the 96 positions in the crystallization tray; Step 2: deposition of nano-liter volumes of this reservoir onto the drop position and deposition of the protein nano-liter volume onto the same spot; Step 3: to refine the condition by making custom grids by varying two components in the lead condition to create new opportunities for improved crystal growth. Another very important feature of the Screenmaker96+8 is that the time required to place the drops (both the precipitant solution and the protein) is significantly less than other crystallization robots, thereby reducing dehydration time by at least half. The crystallization robot will be housed in the newly created Structural Biology Resource Center, supported by The Rockefeller University under the umbrella of the Research Support office. The Center will supervise the robot's use, train users and maintain the instrument in top working condition, and will be responsible for fair, relevant and efficient utilization of the robot. Additional parts needed: computer $2450, installation $4000, two nest chiller system $10450, for sensitive samples and extended warranty $32000. (Prices quoted here do not reflect 10% academic discount applied to total requested in budget, see price quote in Equipment.doc). PUBLIC HEALTH RELEVANCE: Determining the three-dimensional structure of biological macromolecules is key in understanding their function and in opening new venues for designing drugs and cures for human diseases. This can be accomplished through the technique known as X-ray crystallography, where the rate-limiting step is often the crystallization of the target molecule, a menial process that is labor intensive and time consuming. This proposal requests a robot that performs the steps involved in setting up crystallization trials of biologically important molecules, decreasing the labor 10 fold and decreasing the generation of waste while increasing the efficiency and scope of the experimental projects.