NDLI: Variants of Green Fluorescent Protein GFPxm

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Variants of Green Fluorescent Protein GFPxm

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Marine Biotechnology : Volume 8, Issue 4, August 2006

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Variants of Green Fluorescent Protein GFPxm

Content Provider	Springer Nature Link
Author	Luo, Wen Xin Cheng, Tong Guan, Bao Quan Li, Shao Wei Miao, Ji Zhang, Jun Xia, Ning Shao
Copyright Year	2006
Abstract	As research progresses, fluorescent proteins useful for optical marking will evolve toward brighter, monomeric forms that are more diverse in color. We previously reported a new fluorescent protein from Aequorea macrodactyla, GFPxm, that exhibited many characteristics similar to wild-type green fluorescent protein (GFP). However, the application of GFPxm was limited because GFPxm expressed and produced fluorescence only at low temperatures. To improve the fluorescent properties of GFPxm, 12 variants were produced by site-directed mutagenesis and DNA shuffling. Seven of these mutants could produce strong fluorescence when expressed at 37°C. The relative fluorescence intensities of mutants GFPxm16, GFPxm18, and GFPxm19 were higher than that of EGFP (enhanced GFP) when the expression temperature was between 25 and 37°C, and mutants GFPxm16 and GFPxm163 could maintain a high fluorescence intensity even when expressed at 42°C. Meanwhile, at least 4 mutants could be successfully expressed in mammalian cell lines. The fluorescence spectra of 6 of the 12 mutants had a progressive red shift. The longest excitation-emission maximum was at 514/525 nm. In addition, 3 of the 12 mutants had two excitation peaks including an UV-excitation peak, while another mutant had only one UV-excitation peak.
Starting Page	560
Ending Page	566
Page Count	7
File Format	PDF
ISSN	14362228
Journal	Marine Biotechnology
Volume Number	8
Issue Number	5
e-ISSN	14362236
Language	English
Publisher	Springer-Verlag
Publisher Date	2006-07-03
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Fluorescence spectra green fluorescent protein variants Oceanography
Content Type	Text
Resource Type	Article
Subject	Aquatic Science Biotechnology

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