|Ex λ||Em λ||EC (M-1 cm-1)||QY||Brightness||pKa||Maturation (min)||Lifetime (ns)|
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Based on our early results, it seemed as though engineering a true monomer of DsRed might be impossible and therefore we pursued an alternate approach. The basic strategy was to fuse two copies of our best AC dimer with a polypeptide linker such that the critical dimer interactions could be satisfied through intramolecular contacts with the tandem partner encoded within the same polypeptide. With our optimized dimer2 we constructed a series of tandem constructs with linkers of varying lengths (9, 12, 13, or 22 aa) and a sequence similar to a known protease-resistant linker. Of these four, only the tandem construct with the nine-residue linker was notable for its somewhat slower maturation. The other three constructs were practically indistinguishable, and the tandem construct with the 12-residue linker, designated tdimer2(12), is currently our preferred construct because it has the shortest linker.