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Computationally Designed Libraries of Fluorescent Proteins Evaluated by Preservation and Diversity of Function

Thomas P. Treynor, Christina L. Vizcarra, Daniel Nedelcu and Stephen L. Mayo
Proceedings of the National Academy of Sciences of the United States of America
Vol. 104, No. 1 (Jan. 2, 2007), pp. 48-53
Stable URL: http://www.jstor.org/stable/25426048
Page Count: 6
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Computationally Designed Libraries of Fluorescent Proteins Evaluated by Preservation and Diversity of Function
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Abstract

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.

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