GFP‐like chromoproteins as a source of far‐red fluorescent proteins

FEBS Letters

Fradkov, Arkady F.³  Terskikh, Alexey¹  Labas, Yulii A.²  Martynov, Vladimir I.³  Matz, Mikhail V.³  Yanushevich, Yurii G.³  Gurskaya, Nadya G.³  Lukyanov, Sergey A.³  Lukyanov, Konstantin A.³ 

[1] School of Medicine, Stanford University, Stanford, CA 94305, USA;Institute of Ecology and Evolution RAS, Leninsky pr. 33, 117071 Moscow, Russia;Shemiakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, 117997 Moscow, Russia

关键词: Green fluorescent protein;    Anthozoa;    drFP583;    DsRed;    Multi-color labeling;    GFP;    green fluorescent protein;    FP;    fluorescent protein;    CP;    chromoprotein;    cgCP;    Condylactis gigantea chromoprotein;    cpCP;    Condylactis passiflora chromoprotein;    gtCP;    Goniopora tenuidens chromoprotein;    hcCP;    Heteractis crispa chromoprotein;    HcRed;    Heteractis crispa red fluorescent protein;   

DOI  :  10.1016/S0014-5793(01)02930-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 λ _max=615–640 nm). 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.

【 授权许可】

Unknown