Very bright green fluorescent proteins from the Pontellid copepod Pontella mimocerami.
Very bright green fluorescent proteins from the Pontellid copepod Pontella mimocerami.
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DOI：
10.1371/journal.pone.0011517
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发表时间：
2010-07-14
期刊：
影响因子：
3.7
通讯作者：
Matz MV
中科院分区：
文献类型：
--
作者：
Hunt ME;Scherrer MP;Ferrari FD;Matz MV
荧光蛋白（FP）与维多利亚水母（Aequorea victoria）的绿色荧光蛋白（GFP）同源，由于其作为遗传编码的荧光标记的有用性，已经彻底改变了生物医学研究。来自桡足类的荧光蛋白由于其高亮度和快速荧光发展而特别有前途。在这里，我们报告了两个新的FP从Pontella mimocerami（桡足类，哲水蚤，Pontellidae），这是通过荧光筛选的细菌cDNA表达文库制备的动物的全身总RNA。这些蛋白质在序列和光谱性质上非常相似。它们具有高的摩尔消光系数（79，000 M−1 cm−）和量子产率（0.92），这使得它们比最常见的FP标记物EGFP亮两倍多。这两种蛋白质形成寡聚体，我们能够在一定程度上通过N-末端区域的诱变来抵消;然而，这种特定的修饰导致亮度大幅下降。这两种P. mimocerami蛋白的光谱特征使它们成为有史以来最亮的绿色FP。因此，这些蛋白质可能成为体内成像工具包的有价值的补充。
Fluorescent proteins (FP) homologous to the green fluorescent protein (GFP) from the jellyfish Aequorea victoria have revolutionized biomedical research due to their usefulness as genetically encoded fluorescent labels. Fluorescent proteins from copepods are particularly promising due to their high brightness and rapid fluorescence development. Here we report two novel FPs from Pontella mimocerami (Copepoda, Calanoida, Pontellidae), which were identified via fluorescence screening of a bacterial cDNA expression library prepared from the whole-body total RNA of the animal. The proteins are very similar in sequence and spectroscopic properties. They possess high molar extinction coefficients (79,000 M−1 cm−) and quantum yields (0.92), which make them more than two-fold brighter than the most common FP marker, EGFP. Both proteins form oligomers, which we were able to counteract to some extent by mutagenesis of the N-terminal region; however, this particular modification resulted in substantial drop in brightness. The spectroscopic characteristics of the two P. mimocerami proteins place them among the brightest green FPs ever described. These proteins may therefore become valuable additions to the in vivo imaging toolkit.
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影响因子：
3.7
作者：
Alieva NO;Konzen KA;Field SF;Meleshkevitch EA;Hunt ME;Beltran-Ramirez V;Miller DJ;Wiedenmann J;Salih A;Matz MV
通讯作者：
Matz MV
DOI：
10.1073/pnas.97.22.11984
发表时间：
2000-10-24
影响因子：
11.1
作者：
Baird, GS;Zacharias, DA;Tsien, RY
通讯作者：
Tsien, RY
影响因子：
2.7
作者：
AMSTERDAM, A;LIN, S;HOPKINS, N
通讯作者：
HOPKINS, N
影响因子：
56.9
作者：
CHALFIE, M;TU, Y;PRASHER, DC
通讯作者：
PRASHER, DC
影响因子：
1.6
作者：
Deheyn, Dimitri D.;Kubokawa, Kaoru;Holland, Nicholas D.
通讯作者：
Holland, Nicholas D.