GFP-like chromoproteins as a source of far-red fluorescent proteins
GFP-like chromoproteins as a source of far-red fluorescent proteins
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DOI：
10.1016/s0014-5793(01)02930-1
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发表时间：
2001-10-19
期刊：
影响因子：
3.5
通讯作者：
Lukyanov, SA
中科院分区：
文献类型：
--
作者：
Gurskaya, NG;Fradkov, AF;Lukyanov, SA
我们采用了一种新的方法来从红色吸收色素蛋白中制备新型荧光蛋白(FP)。一种相同的单一氨基酸替代将来自珊瑚虫物种的新的色素蛋白(Heteractis crispa，Condylactis Gigantea，和Goniopora tenuidens)转化为远红FFP(发射波长(Max)=615-640mN)。此外，结合定点突变和随机突变，获得了一种独特的远红光荧光蛋白(HcRed)，该荧光蛋白在645 nm处有明亮的发射。与drFP583相比，HcRed的荧光发生了明显的红移(超过60 nm)，使其成为多色标记的理想附加颜色。重要的是，HcRed可被600 nm染料激光激发，从而为多色流式细胞术应用提供了新的检测通道。此外，我们还产生了一个二聚体突变体，它的成熟度和光谱性质与四聚体HcRed相似。(C)2001年，埃尔塞维尔科学公司代表欧洲生物化学学会联合会出版。
We have employed a new approach to generate novel fluorescent proteins (FPs) from red absorbing chromoproteins. An identical single amino acid substitution converted novel chromoproteins from the species Anthozoa (Heteractis crispa, Condylactis gigantea, and Goniopora tenuidens) into far-red FPs (emission lambda (max) = 615-640 mn). Moreover, coupled site-directed and random mutagenesis of the chromoprotein from H. crispa resulted in a unique far-red FP (HcRed) that exhibited bright emission at 645 nm. A clear red shift in fluorescence of HcRed, compared to drFP583 (by more than 60 nm), makes it an ideal additional color for multi-color labeling. Importantly, HcRed is excitable by 600 nm dye laser, thus promoting new detection channels for multi-color flow cytometry applications. In addition, we generated a dimeric mutant with similar maturation and spectral properties to tetrameric HcRed. (C) 2001 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.