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Figure 1.

Evolution of the subfamily of orange fluorescent proteins.

(A) Phylogenic tree showing the history of the development of different orange fluorescent proteins. (B) Chemical structures of the chromophores found in orange fluorescent proteins.

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Table 1.

Spectral and evolutionary features of orange fluorescent proteins.

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Table 2.

Data collection and refinement statistics.

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Figure 2.

Amino acid alignment of mOrange, LSSmOrange, PSmOrange and PSmOrange2.

The chromophore-forming tri-peptides are highlighted in yellow.

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Figure 3.

Amino acid differences between the parental and successor proteins in 3D.

(A) The transformation of mOrange into LSSmOrange. (B) The transformation of mOrange into PSmOrange. (C) The transformation of PSmOrange into PSmOrange2.

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Figure 4.

The differences in the immediate chromophore environment between the parental and successor proteins.

(A) The difference between mOrange and LSSmOrange. (B) The difference between mOrange and PSmOrange. (C) The difference between PSmOrange and PSmOrange2.

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Figure 5.

The structures of the PSmOrange and DsRed chromophores.

(A) The structure of the orange form of the PSmOrange chromophore. (B) The modeled structure of the photoconverted far-red form of the PSmOrange chromophore. (C) The structure of the DsRed chromophore.

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Figure 6.

An evolution of orange fluorescent proteins derived from KO, DsRed and zFP538 with mutations critical to the phenotype of each variant.

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Figure 7.

Possibilities for the future design of advanced orange and far-red fluorescent proteins.

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