Mammalian expression of infrared fluorescent proteins engineered from a bacterial phytochrome.
Mammalian expression of infrared fluorescent proteins engineered from a bacterial phytochrome.
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DOI：
10.1126/science.1168683
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发表时间：
2009-05-08
期刊：
影响因子：
--
通讯作者：
Tsien RY
中科院分区：
文献类型：
--
作者：
Shu X;Royant A;Lin MZ;Aguilera TA;Lev-Ram V;Steinbach PA;Tsien RY
来自水母和珊瑚的可见荧光蛋白（FP）已经彻底改变了分子和细胞生物学的许多领域，但是FP在小鼠等完整动物中的使用受到激发光穿透性差的阻碍。我们现在证明，从耐辐射球菌中提取的细菌光敏色素，结合胆绿素作为发色团，可以被改造成单体的红外荧光蛋白（IFPs），其激发/发射最大值分别为684/708 nm，消光系数> 90，000 M−1cm−1，量子产率为0.07。IFPs在哺乳动物细胞和小鼠中表达良好，并自发掺入胆绿素，胆绿素作为血红素催化剂的初始中间体普遍存在，但本身具有可忽略的荧光。由于其波长能很好地穿透组织，IFPs适用于全身成像。这里开发的IFPs为进一步的工程设计提供了一个支架。
Visibly fluorescent proteins (FPs) from jellyfish and corals have revolutionized many areas of molecular and cell biology, but the use of FPs in intact animals such as mice has been handicapped by poor penetration of excitation light. We now show that a bacteriophytochrome from Deinococcus radiodurans incorporating biliverdin as the chromophore can be engineered into monomeric, infrared-fluorescent proteins (IFPs), with excitation/emission maxima of 684/708 nm respectively, extinction coefficient > 90,000 M−1cm−1, and quantum yield 0.07. IFPs express well in mammalian cells and mice, and spontaneously incorporate biliverdin, which is ubiquitous as the initial intermediate in heme catabolism but has negligible fluorescence by itself. Because their wavelengths penetrate tissue well, IFPs are suitable for whole-body imaging. The IFPs developed here provide a scaffold for further engineering.
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DOI：
10.1073/pnas.0407645101
发表时间：
2004-12-14
影响因子：
11.1
作者：
Fischer, AJ;Lagarias, JC
通讯作者：
Lagarias, JC
DOI：
10.1073/pnas.0806718105
发表时间：
2008-09-23
影响因子：
11.1
作者：
Yang, Xiaojing;Kuk, Jane;Moffat, Keith
通讯作者：
Moffat, Keith
影响因子：
9.8
作者：
Rusch DB;Halpern AL;Sutton G;Heidelberg KB;Williamson S;Yooseph S;Wu D;Eisen JA;Hoffman JM;Remington K;Beeson K;Tran B;Smith H;Baden-Tillson H;Stewart C;Thorpe J;Freeman J;Andrews-Pfannkoch C;Venter JE;Li K;Kravitz S;Heidelberg JF;Utterback T;Rogers YH;Falcón LI;Souza V;Bonilla-Rosso G;Eguiarte LE;Karl DM;Sathyendranath S;Platt T;Bermingham E;Gallardo V;Tamayo-Castillo G;Ferrari MR;Strausberg RL;Nealson K;Friedman R;Frazier M;Venter JC
通讯作者：
Venter JC
影响因子：
29.4
作者：
Nakao, A;Otterbein, LE;Bach, FH
通讯作者：
Bach, FH
影响因子：
56.9
作者：
Davis, SJ;Vener, AV;Vierstra, RD
通讯作者：
Vierstra, RD