红色荧光蛋白mCherry中插入外源短肽位点的研究

生物技术通报 ›› 2013, Vol. 0 ›› Issue (5): 144-148.

红色荧光蛋白mCherry中插入外源短肽位点的研究

梁俊婷^{1, 2}, 李鹿之², 陈少鹏², 焦浈¹

(1.郑州大学离子束生物工程省重点实验室，郑州 450052；2.中国科学院合肥物质科学研究院技术生物与农业工程研究所，合肥 230031)

收稿日期:2012-11-12 修回日期:2013-05-24 出版日期:2013-05-24 发布日期:2013-05-24
作者简介:梁俊婷，女，硕士研究生，研究方向：生物大分子的人工定向进化；E-mail：lynne5601@hotmail.com

Study on Sites of the Tolerate Peptide Insertion in the Fluorescent Protein of mCherry

Liang Junting^{1, 2}, Li Luzhi², Chen Shaopeng², Jiao Zhen¹

(1. Henan Provincial Key Laboratory of Ion Beam Bioengineering，Zhengzhou University，Zhengzhou 450052；2. Institute of Technical Biology & Agriculture Engineering，Chinese Academy of Sciences，Hefei 230031)

Received:2012-11-12 Revised:2013-05-24 Published:2013-05-24 Online:2013-05-24
About author:焦浈，女，博士，副教授，研究方向：作物遗传育种；E-mail：jiaozhen@zzu.edu.cn

摘要/Abstract

摘要： 荧光蛋白及其突变体广泛应用于细胞生物学、分子生物学及医学等领域，而围绕荧光蛋白建立的各种新技术，如BiFC(双分子荧光互补技术)、FRET(荧光能量共振转移技术)、Optical highlighter(荧光笔探针)等，则极大地促进了多个生物相关学科的发展。采用转座子突变技术，在红色荧光蛋白mCherry中随机插入5个氨基酸残基。经流式细胞仪检测和分选，成功获得13个插入外源氨基酸短肽后，仍具有荧光活性的mCherry荧光蛋白突变体，为BiFC以及构建新型生物感受器提供了更多可插入外源DNA的合适位点。

关键词: 红色荧光蛋白, 短肽, 插入位点

Abstract: The fluorescent proteins(FPs)and their color variants have been widely used in cell biology, molecular biology and medicine. Various new technologies based on these proteins such as bimolecular fluorescence complementation (BiFC), fluorescence resonance energy transfer (FRET)-based techniques and optical highlighter are enlightening the whole biology technology. In this paper, we used transposon-based mutagenesis and flow cytometry to verify thirteen tolerate sites of mCherry with five amino acids inserted which fluoresced red. The inserted variants with fluorescence can be served for BiFC and provide more selective sites for designing of bio-sensors.

Key words: Red fluorescent protein, Short-peptide, Inserted sites

梁俊婷, 李鹿之, 陈少鹏, 焦浈. 红色荧光蛋白mCherry中插入外源短肽位点的研究[J]. 生物技术通报, 2013, 0(5): 144-148.

Liang Junting, Li Luzhi, Chen Shaopeng, Jiao Zhen. Study on Sites of the Tolerate Peptide Insertion in the Fluorescent Protein of mCherry[J]. Biotechnology Bulletin, 2013, 0(5): 144-148.

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