Computationally designed libraries of fluorescent proteins evaluated by preservation and diversity of function
Computationally designed libraries of fluorescent proteins evaluated by preservation and diversity of function
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DOI：
10.1073/pnas.0609647103
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发表时间：
2007-01-02
影响因子：
11.1
通讯作者：
Mayo, Stephen L.
中科院分区：
文献类型：
--
作者：
Treynor, Thomas P.;Vizcarra, Christina L.;Mayo, Stephen L.
为了确定七种文库设计算法中哪一种最好地引入新的蛋白质功能而不完全破坏它，设计并合成了七种绿色荧光蛋白变体的组合文库。每种方法都通过体内测量所得的发射强度和颜色分布进行评估。与通过易错PCR构建的文库进行另外的比较。在所设计的库中，荧光功能被保存在通过使用本文开发和描述的基于结构的计算方法设计的库中的样品的最大部分。在更好地保留荧光的设计文库中观察到朝向更大的颜色多样性的趋势。与通过易错PCR构建的文库中观察到的趋势相反，在用基于结构的计算方法设计的四个文库中，观察到功能的保留随着文库的平均突变水平而增加。
To determine which of seven library design algorithms best introduces new protein function without destroying it altogether, seven combinatorial libraries of green fluorescent protein variants were designed and synthesized. Each was evaluated by distributions of emission intensity and color compiled from measurements made in vivo. Additional comparisons were made with a library constructed by error-prone PCR. Among the designed libraries, fluorescent function was preserved for the greatest fraction of samples in a library designed by using a structure-based computational method developed and described here. A trend was observed toward greater diversity of color in designed libraries that better preserved fluorescence. Contrary to trends observed among libraries constructed by error-prone PCR, preservation of function was observed to increase with a library's average mutation level among the four libraries designed with structure-based computational methods.