Modular generation of fluorescent phycobiliproteins
Modular generation of fluorescent phycobiliproteins
复制标题
荧光藻胆蛋白的模块化生成
DOI：
10.1039/c3pp25383j
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发表时间：
2013-01-01
影响因子：
3.1
通讯作者：
Zhao, Kai-Hong
中科院分区：
文献类型：
--
作者：
Wu, Xian-Jun;Chang, Kun;Zhao, Kai-Hong
藻胆蛋白是一种明亮的荧光捕光色素，用于蓝藻和红藻的光合作用。它们作为荧光生物标志物也很有趣，但它们在体内的异源产生以前需要多次转化。我们在这里报告了一种只需要两个 DNA 片段的模块化方法。第一个代码为脱辅基蛋白。第二个编码能够进行发色团生物合成及其与脱辅基蛋白共价连接的融合体；它含有血红素加氧酶、胆素还原酶和发色团裂解酶的基因。在大肠杆菌中高产地获得了含有藻红蛋白（λ _flor ~ 560 nm）、藻胆蛋白（λ _flor ~ 500 nm）、藻蓝蛋白（λ _flor ~ 630 nm）或藻紫红蛋白（λ _flor ~ 580 nm）的藻胆蛋白。这种方法有利于藻胆蛋白的发色研究，以及基于其高荧光的荧光标记的使用。
Phycobiliproteins are brightly-fluorescent light-harvesting pigments for photosynthesis in cyanobacteria and red algae. They are also of interest as fluorescent biomarkers, but their heterologous generation in vivo has previously required multiple transformations. We report here a modular approach that requires only two DNA segments. The first codes for the apo-protein. The second codes for fusions capable of chromophore biosynthesis and its covalent attachment to the apo-protein; it contains the genes of heme oxygenase, a bilin reductase, and a chromophore lyase. Phycobiliproteins containing phycoerythrobilin ( λ _fluor ~ 560 nm), phycourobilin ( λ _fluor ~ 500 nm), phycocyanobilin ( λ _fluor ~ 630 nm) or phycoviolobilin ( λ _fluor ~ 580 nm) were obtained in high yield in E. coli . This approach facilitates chromophorylation studies of phycobiliproteins, as well as their use for fluorescence labeling based on their high fluorescence.