Towards rationalization of crystallization screening for small- to medium-sized academic laboratories: the PACT/JCSG+ strategy.
Towards rationalization of crystallization screening for small- to medium-sized academic laboratories: the PACT/JCSG+ strategy.
复制标题
实现中小型学术实验室结晶筛选的合理化：PACT/JCSG 策略。
DOI：
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发表时间：
2005
期刊：
影响因子：
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通讯作者：
A. Perrakis
中科院分区：
文献类型：
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作者：
J. Newman;D. Egan;T. Walter;Ran Meged;I. Berry;Marouane Ben Jelloul;J. Sussman;D. Stuart;A. Perrakis
介绍了为小型学术实验室开发的结晶筛选工艺。其基本概念是在最少数量的结晶条件下将联合收割机稀疏矩阵筛选与系统筛选相结合。稀疏矩阵筛选是从结构基因组学联合中心（JCSG）中挑选的条件组合，使用来自其他筛选的条件进行扩展。其目的是最大限度地覆盖结晶参数空间，没有冗余。系统筛选，pH值，阴离子和阳离子测试（PACT）屏幕，旨在解耦的组件，每种条件，并提供有关蛋白质的信息，即使在没有晶体，而不是覆盖广泛的结晶空间。这种筛选策略与纳升体积的点胶硬件和一个小而实用的实验跟踪系统相结合。筛选已在NKI和其他实验室进行了测试，并得出结论，他们提供了一个有用的最小筛选策略。
A crystallization screening process is presented that was developed for a small academic laboratory. Its underlying concept is to combine sparse-matrix screening with systematic screening in a minimum number of crystallization conditions. The sparse-matrix screen is the cherry-picked combination of conditions from the Joint Center for Structural Genomics (JCSG) extended using conditions from other screens. Its aim is to maximize the coverage of crystallization parameter space with no redundancy. The systematic screen, a pH-, anion- and cation-testing (PACT) screen, aims to decouple the components of each condition and to provide information about the protein, even in the absence of crystals, rather than cover a wide crystallization space. This screening strategy is combined with nanolitre-volume dispensing hardware and a small but practical experiment-tracking system. The screens have been tested both at the NKI and in other laboratories and it is concluded that they provide a useful minimal screening strategy.