GFP研究界的權(quán)威---錢(qián)永佑（美國(guó)科學(xué)院院士）,。Roger Tsien大家可能并不熟,，但一提起GFP,現(xiàn)在是無(wú)人不曉,，也算是一項(xiàng)很有意義的工作，對(duì)生物學(xué)的研究具有不小的推動(dòng),。Roger Tsien的實(shí)驗(yàn)室是最早用GFP來(lái)標(biāo)記protein的,，并不斷開(kāi)發(fā)GFP的應(yīng)用。例如,，大部分GFP的突變,，YFP（yellow),CFP(cyan)，BFP（blue)
等等,。據(jù)稱(chēng)有望獲得諾貝爾獎(jiǎng)的華人,。

個(gè)人的CV如下：

 
Roger Y. Tsien, Ph.D.
Investigator,
University of California, San Diego
 
GFP-Based Indicators of Redox Status
Colette Dooley has been exploiting and modifying indicators of cellular oxidation-reduction status first created by George Hanson in the laboratory of S. James Remington (University of Oregon). These molecules are mutants of green fluorescent protein (GFP) to which pairs of cysteines have been added. Under oxidizing conditions, these cysteines form intramolecular disulfide bonds, which distort the chromophore inside the GFP and thus change its fluorescence reversibly. Dooley extensively characterized these molecules both in free solution and inside cells. In the cytosol of healthy cells, the indicators report a redox potential of about –320 mV, considerably more reducing than previously determined by traditional destructive biochemical assays. In leukocyte-like cell lines stimulated to generate superoxide and hydrogen peroxide, the indicators become substantially oxidized, showing that they are capable of responding to endogenous bursts of reactive oxygen species. However, stimulation by growth factors, which has also been reported to generate similar species, has consistently failed to give any detectable signals, despite many physiological manipulations or three types of mutations that made the indicators more sensitive to exogenous hydrogen peroxide. These negative results are leading us to question whether growth factors really cause significant production of reactive oxygen species.

Red Fluorescent Proteins
Nathan Shaner, an HHMI predoctoral scholar, continued work started by Robert Campbell to improve engineered dimeric and monomeric red fluorescent proteins. Our previous best mutant, mRFP1, has been supplied to more than 1,000 requesting laboratories but still has imperfections, such as a significant fraction of protein that never becomes fluorescent. Continued evolution has increased the range of colors, the fraction of correct maturation, and the quantum yield of fluorescence of some of the colors. Five distinguishable hues from yellow to far red, with emission peaks at 558, 569, 581, 612, and 632 nm now exist, though no one molecule is optimal in all respects.
文章（大多免費(fèi)閱讀，請(qǐng)點(diǎn)擊文章的作者即可打開(kāi)pubmed,并找到原文）

1:  Dooley CM, Dore TM, Hanson GT, Jackson WC, Remington SJ, Tsien RY. 
 Imaging dynamic redox changes in mammalian cells with green fluorescent protein indicators.
J Biol Chem. 2004 Feb 25
 

2 Tsien RY. Imagining imaging's future.
Nat Rev Mol Cell Biol. 2003 Sep;Suppl:SS16-21. Review.

3 Zhang J, Campbell RE, Ting AY, Tsien RY. Creating new fluorescent probes for cell biology.
Nat Rev Mol Cell Biol. 2002 Dec;3(12):906-18. Review.

4 Campbell RE, Tour O, Palmer AE, Steinbach PA, Baird GS, Zacharias DA, Tsien RY. A monomeric red fluorescent protein.Proc Natl Acad Sci U S A. 2002 Jun 11;99(12):7877-82.
5 Zacharias DA, Violin JD, Newton AC, Tsien RY. Partitioning of lipid-modified monomeric GFPs into membrane microdomains of live cells.Science. 2002 May 3;296(5569):913-6.

6 Siegel RM, Chan FK, Zacharias DA, Swofford R, Holmes KL, Tsien RY, Lenardo MJ. Measurement of molecular interactions in living cells by fluorescence resonance energy transfer between variants of the green fluorescent protein. Sci STKE. 2000 Jun 27;2000(38):PL1.

7 Ting AY, Kain KH, Klemke RL, Tsien RY. Genetically encoded fluorescent reporters of protein tyrosine kinase activities in living cells.
Proc Natl Acad Sci U S A. 2001 Dec 18;98(26):15003-8.

8 Tsien RY, Bacskai BJ, Adams SR. FRET for studying intracellular signalling.
Trends Cell Biol. 1993 Jul;3(7):242-5. No abstract available.

9 Hanson GT, Aggeler R, Oglesbe D, Canon M, Capaldi RA, Tsien RY, Remington SJ. Investigating mitochondrial redox potential with redox-sensitive green fluorescent protein indicators.
J Biol Chem. 2004 Jan 13