We have cloned an unusual colourless green fluorescent protein (GFP)-like protein from Aequorea coerulescens (acGFPL). The A. coerulescens specimens displayed blue (not green) luminescence, and no fluorescence was detected in these medusae. Escherichia coli expressing wild-type acGFPL showed neither fluorescence nor visible coloration. Random mutagenesis generated green fluorescent mutants of acGFPL, with the strongest emitters found to contain an Glu222→Gly (E222G) substitution, which removed the evolutionarily invariant Glu222. Re-introduction of Glu222 into the most fluorescent random mutant, named aceGFP, converted it into a colourless protein. This colourless aceGFP-G222E protein demonstrated a novel type of UV-induced photoconversion, from an immature non-fluorescent form into a green fluorescent form. Fluorescent aceGFP may be a useful biological tool, as it was able to be expressed in a number of mammalian cell lines. Furthermore, expression of a fusion protein of ‘humanized’ aceGFP and β-actin produced a fluorescent pattern consistent with actin distribution in mammalian cells.
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July 2003
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Research Article|
July 15 2003
A colourless green fluorescent protein homologue from the non-fluorescent hydromedusa Aequorea coerulescens and its fluorescent mutants
Nadya G. GURSKAYA
;
Nadya G. GURSKAYA
∗Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, Moscow 117997, Russia
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Arkady F. FRADKOV
;
Arkady F. FRADKOV
∗Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, Moscow 117997, Russia
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Natalia I. POUNKOVA
;
Natalia I. POUNKOVA
∗Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, Moscow 117997, Russia
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Dmitry B. STAROVEROV
;
Dmitry B. STAROVEROV
†Evrogen JSC, Miklukho-Maklaya 16/10, Moscow 117997, Russia
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Maria E. BULINA
;
Maria E. BULINA
∗Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, Moscow 117997, Russia
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Yurii G. YANUSHEVICH
;
Yurii G. YANUSHEVICH
∗Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, Moscow 117997, Russia
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Yulii A. LABAS
;
Yulii A. LABAS
‡Institute of Biochemistry RAS, Leninsky pr. 33, Moscow 117071, Russia
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Sergey LUKYANOV
;
Sergey LUKYANOV
∗Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, Moscow 117997, Russia
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Konstantin A. LUKYANOV
Konstantin A. LUKYANOV
1
∗Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, Moscow 117997, Russia
1To whom correspondence should be addressed (e-mail kluk@ibch.ru).
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Biochem J (2003) 373 (2): 403–408.
Article history
Received:
December 19 2002
Revision Received:
March 04 2003
Accepted:
April 14 2003
Accepted Manuscript online:
April 14 2003
Citation
Nadya G. GURSKAYA, Arkady F. FRADKOV, Natalia I. POUNKOVA, Dmitry B. STAROVEROV, Maria E. BULINA, Yurii G. YANUSHEVICH, Yulii A. LABAS, Sergey LUKYANOV, Konstantin A. LUKYANOV; A colourless green fluorescent protein homologue from the non-fluorescent hydromedusa Aequorea coerulescens and its fluorescent mutants. Biochem J 15 July 2003; 373 (2): 403–408. doi: https://doi.org/10.1042/bj20021966
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