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连续C末端色氨酸对绿色荧光蛋白荧光的影响。

Effect of sequential C-terminal tryptophans on green fluorescent protein fluorescence.

作者信息

Tsuchida Shirou, Kanashiki Takumi, Izumiya Shuhei, Ichikawa Takuya, Kurosawa Ryusuke, Hamaue Naoya, Aoki Takashi

机构信息

Department of Molecular Biosciences School of Pharmaceutical Sciences Health Sciences University of Hokkaido Ishikari-Tobetsu Japan.

出版信息

FEBS Open Bio. 2018 May 29;8(7):1176-1183. doi: 10.1002/2211-5463.12445. eCollection 2018 Jul.

DOI:10.1002/2211-5463.12445
PMID:29988552
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6026694/
Abstract

The effect of the addition of sequential C-terminal tryptophan residues on the fluorescence intensity of GFP was investigated. Tandem repeats of six tryptophan residues markedly decreased fluorescence intensity. This phenomenon is likely to occur because of the inhibition of GFP folding, resulting in insolubility. Exploiting this phenomenon, we constructed a cloning vector that facilitates the identification of recombinant colonies of by the activation of GFP.

摘要

研究了连续添加C端色氨酸残基对绿色荧光蛋白（GFP）荧光强度的影响。六个色氨酸残基的串联重复显著降低了荧光强度。这种现象可能是由于GFP折叠受到抑制，导致不溶性。利用这一现象，我们构建了一种克隆载体，通过激活GFP来促进重组菌落的鉴定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/12b6/6026694/2b0926b106a3/FEB4-8-1176-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/12b6/6026694/f64e03cbb841/FEB4-8-1176-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/12b6/6026694/dc6cc4078854/FEB4-8-1176-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/12b6/6026694/df5eaa02f46e/FEB4-8-1176-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/12b6/6026694/2b0926b106a3/FEB4-8-1176-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/12b6/6026694/f64e03cbb841/FEB4-8-1176-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/12b6/6026694/dc6cc4078854/FEB4-8-1176-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/12b6/6026694/df5eaa02f46e/FEB4-8-1176-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/12b6/6026694/2b0926b106a3/FEB4-8-1176-g004.jpg

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Effect of sequential C-terminal tryptophans on green fluorescent protein fluorescence.连续C末端色氨酸对绿色荧光蛋白荧光的影响。
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本文引用的文献

1
Screening of recombinant Escherichia coli using activation of green fluorescent protein as an indicator.以绿色荧光蛋白激活作为指标筛选重组大肠杆菌。
Biochem Biophys Res Commun. 2014 Sep 12;452(1):32-5. doi: 10.1016/j.bbrc.2014.08.038. Epub 2014 Aug 17.
2
Subcellular localization of transiently expressed fluorescent fusion proteins.瞬时表达的荧光融合蛋白的亚细胞定位
Methods Mol Biol. 2013;1069:227-58. doi: 10.1007/978-1-62703-613-9_16.
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NIH Image to ImageJ: 25 years of image analysis.NIH 图像到 ImageJ：25 年的图像分析。
Nat Methods. 2012 Jul;9(7):671-5. doi: 10.1038/nmeth.2089.
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Imaging molecular pathways: reporter genes.成像分子途径：报告基因。
Radiat Res. 2012 Apr;177(4):508-13. doi: 10.1667/rr2918.1. Epub 2012 Feb 21.
5
Fluorescence resonance energy transfer-based assay for DNA-binding protein tagged by green fluorescent protein.基于荧光共振能量转移的绿色荧光蛋白标记的DNA结合蛋白检测方法。
Biosci Biotechnol Biochem. 2006 Aug;70(8):1921-7. doi: 10.1271/bbb.60085.
6
Engineering and characterization of a superfolder green fluorescent protein.超折叠绿色荧光蛋白的工程设计与特性研究
Nat Biotechnol. 2006 Jan;24(1):79-88. doi: 10.1038/nbt1172. Epub 2005 Dec 20.
7
Protein tagging and detection with engineered self-assembling fragments of green fluorescent protein.利用绿色荧光蛋白的工程自组装片段进行蛋白质标记与检测。
Nat Biotechnol. 2005 Jan;23(1):102-7. doi: 10.1038/nbt1044. Epub 2004 Dec 5.
8
His tag effect on solubility of human proteins produced in Escherichia coli: a comparison between four expression vectors.他的标签对大肠杆菌中产生的人类蛋白质溶解度的影响：四种表达载体之间的比较。
J Struct Funct Genomics. 2004;5(3):217-29. doi: 10.1023/b:jsfg.0000031965.37625.0e.
9
Extraction, purification and properties of aequorin, a bioluminescent protein from the luminous hydromedusan, Aequorea.来自发光水螅水母（海月水母属）的生物发光蛋白水母发光蛋白的提取、纯化及特性
J Cell Comp Physiol. 1962 Jun;59:223-39. doi: 10.1002/jcp.1030590302.
10
General properties of GFP-display, an electrophoretic analysis for single amino acid changes in target polypeptides.绿色荧光蛋白展示的一般特性，一种用于分析目标多肽中单个氨基酸变化的电泳分析方法。
Anal Biochem. 2003 Jun 1;317(1):107-15. doi: 10.1016/s0003-2697(03)00112-x.