环化荧光蛋白生物探针荧光寿命成像研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2018-0212

摘要/Abstract

摘要： 环化荧光蛋白探针作为一种新型的基因编码生物探针,是由荧光蛋白和对探测物有特异性响应的结合域蛋白构成的。由于其独特的构造方式使其在对探测物检测时具有较高的灵敏度和较大的动态范围,引起了越来越多科学家的兴趣,已发展成为监测生命活动的一种强有力工具。目前越来越多基于环化荧光蛋白技术开发的生物探针被广泛的应用于生物成像、药物筛选、细胞代谢物检测等领域,并发挥出了越来越大的作用。但是,这类探针目前仍然存在着开发工作量大,部分发光强度较弱及测量方法受限等问题,解决这些问题还需要科学家们不断的努力。对环化荧光蛋白生物探针进行综述,主要介绍了环化荧光蛋白生物探针的设计方法,已有探针的种类和基本性质,现阶段的发展状况以及存在的问题。另外,介绍了多种检测方法在环化荧光蛋白生物探针上面的使用,特别是近期出现的荧光寿命成像技术在其应用上的进展,比较了这些检测方法的优劣,旨在从多方面阐述环化荧光蛋白生物探针的发展现状,以便能给相关研究者提供一些参考和启发。

关键词: 生物探针, 环化荧光蛋白, 荧光寿命成像

Abstract: Circularly-permuted fluorescent protein（cpFP）biosensors as novel genetically encoded biosensors consist of fluorescent protein and binding domain protein that specifically responds to typical analyte. There is relatively high sensitivity and large dynamic range during the detection of analyte duo to cpFP biosensor’ unique mode of construction,which have attracted more and more scientists’ interests,thus they have become powerful tools for monitoring life activities. Nowadays,growingly biosensors based on cpFP have been widely used in the fields of biological imaging,drug screening and cellular metabolites detection,i.e.,played an increasingly important role. However,there are issues such as development workload is very heavy,fluorescence intensity is weak,and measurement method is limited while developing these sensors. Constant efforts from scientists are still highly required to solve these issues. In this paper,we reviewed researches on the cpFP biosensors,focusing on the design method of cpFP biosensors,the types and basic properties of the current cpFP biosensors,the current states of development,and existing problems. In addition,we introduced the detection methods used in cpFP biosensors especially the fluorescence lifetime imaging method,and compared the advantages and disadvantages of these detection methods. This article aims to elaborate the development status of cpFP biosensors from various aspects,so as to provide researchers references and inspirations.

Key words: biosensor, circularly-permuted fluorescent protein, fluorescence lifetime imaging

周加胜, 王鹏, 周黄梅, 许金铭, 张三军. 环化荧光蛋白生物探针荧光寿命成像研究进展[J]. 生物技术通报, 2018, 34(10): 71-80.

ZHOU Jia-sheng, WANG Peng, ZHOU Huang-mei, XU Jin-ming, ZHANG San-jun. Progress in Fluorescence Lifetime Imaging of Circularly-permuted Fluorescent Protein Biosensors[J]. Biotechnology Bulletin, 2018, 34(10): 71-80.

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