CORE--RNA SYNTHESIS AND RECOMBINANT PROTEIN EXPRESSION FACILITY

RNA合成和重组蛋白核心是一个共享的设施 将提供试剂以促进拟议的研究； 重组蛋白生产设施将提供试剂。按顺序 为了方便实验建议，毫克量的纯 包含HIV-1基因组RT-U5-PBS区域和 将需要合成的tRNA。为了提供这项服务，核心将 重组T7依赖DNA的RNA聚合酶(T7)的表达和纯化 聚合酶)，纯化重组质粒，并进行体外转基因 剧本。一种寡核苷酸合成器被要求生成 经过特别修饰的RNA、DNA模板和引物。核心意志 提供重组逆转录酶(RT)和核衣壳(NCp7) 蛋白质。将这些活动集中在共享资源中 复合体将使这些试剂能够以高效和 成本效益的方式，并将利用现有的试验 这方面的专业知识。

The RNA Synthesis and Recombinant Protein Core is a shared facility which will provide reagents to facilitate the studies proposed; the Recombinant Protein Production Facility will provide reagents. In order to facilitate the experiments proposed, milligram amounts of pure recombinant RNA encompassing the RT-U5-PBS region of HIV-1 genomes and synthetic tRNA will be needed. To provide this service, the Core will express and purify recombinant T7 DNA dependent RNA polymerase (T7 polymerase), purify recombinant plasmids and perform in vitro tran- scriptions. An oligonucleotide synthesizer is requested to generate specifically modified RNAs and DNA templates and primers. The Core will provide recombinant reverse transcriptase (RT) and nucleocapsid (NCp7) proteins. The concentration of these activities in a shared resource complex will enable these reagents to be derived in an efficient and cost effect manner and will capitalize on the available experimental expertise in this area.