基于基因密码子扩展技术的protrudin蛋白定点荧光标记
首发时间:2014-11-26
摘要:目的:将荧光分子引入到细胞内protrudin蛋白特异位点,从而在活体实现对蛋白的动态可视化标记。方法:利用基因密码子扩展技术,将非天然氨基酸(Lys-N3)定点插入到protrudin蛋白中,并通过无铜点击化学引入荧光分子。结果:DIBO-Alexa fluor-555与protrudin蛋白上Lys-N3的click反应可以进行活细胞染色,同时进行免疫荧光染色,发现两种对protrudin染色的方法有共定位趋势。结论:采用新方法实现了活细胞内protrudin特异位点的荧光标记,进而可以示踪其在细胞内的分布及转运过程。?????
关键词: 蛋白质标记 protrudin 点击化学 非天然氨基酸?????
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Site-specific fluorescence labeling of protrudin based on "genetic code expansion technology"
Abstract:Objective:to introduce fluorescent probe into protrudin in living cells so as to achieve the dynamic visulization of protein in vivo. Methods:incorporate unnatural amino acid, Lys-N3 into key sites of protrudin and link DIBO-Alexa fluor-555 by click chemistry, then observe the staining of protrudin in Hela cells. Results: the staining by the new method of fluorescence labeling of protrudin can overlay with the traditional method of immunofluorescence. Conclusions: the site-specifically labeling of protrudin in living cells was achieved and can be used to observe the distribution and translocation of protrudin in vivo.
Keywords: protein labeling protrudin click chemistry: unnatural amino acid
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No.4619389101588314****
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