Finley James B, Qiu Shi-Hong, Luan Chi-Hao, Luo Ming
Center for Biophysical Sciences and Engineering, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Protein Expr Purif. 2004 Mar;34(1):49-55. doi: 10.1016/j.pep.2003.11.026.
The structural genomics initiatives have begun with the aim to create a so-called "basic set library" of protein folds that will be used to improve protein prediction methods. Such a library is thought to require the determination of up to 10,000 new structures, including representative structures of several sequence variants from each protein fold. To meet this goal in a reasonable time frame and cost, automated systems must be utilized to clone and to identify the soluble recombinant proteins contained in multiple genomes. This paper presents such a system, developed using the genome of Caenorhabditis elegans (19,099 genes) as a model eukaryotic organism for structural genomics. This system successfully automates nearly all aspects of recombinant protein expression analysis including subcloning, bacterial growth, recombinant protein expression, protein purification, and scoring protein solubility.
结构基因组学计划已经启动，旨在创建一个所谓的蛋白质折叠“基本集文库”，该文库将用于改进蛋白质预测方法。这样一个文库被认为需要测定多达10000个新结构，包括来自每个蛋白质折叠的几个序列变体的代表性结构。为了在合理的时间框架和成本内实现这一目标，必须利用自动化系统来克隆和鉴定多个基因组中包含的可溶性重组蛋白。本文介绍了这样一个系统，它是以秀丽隐杆线虫基因组（19099个基因）作为结构基因组学的模型真核生物开发的。该系统成功地将重组蛋白表达分析的几乎所有方面自动化，包括亚克隆、细菌生长、重组蛋白表达、蛋白纯化以及评估蛋白溶解性。
