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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Borisova V.V., Frank L.A., Markova S.V., Burakova L.P., Vysotski E.S.
Заглавие : Recombinant Metridia luciferase isoforms: expression, refolding and applicability for in vitro assay
Колич.характеристики :7 с
Коллективы :
Место публикации : Photochem. Photobiol. Sci.: ROYAL SOC CHEMISTRY, 2008. - Vol. 7, Is. 9. - С. 1025-1031. - ISSN 1474-905X, DOI 10.1039/b807271j
Примечания : Cited References: 19. - The work was supported by Bayer AG, by the Russian Foundation for Basic Research grants 05-04-48271 and 06-04-08076, by the joint grant 06-04-89502 of the Russian Foundation for Basic Research and Taiwan National Science Council, and by the "Molecular and Cellular Biology" program of the Russian Academy of Sciences.
Предметные рубрики: BIOLUMINESCENT REPORTER
GAUSSIA LUCIFERASE
CDNA
PROTEINS
CLONING
OVEREXPRESSION
PURIFICATION
MUTAGENESIS
ENZYME
OBELIN
Аннотация: The recombinant coelenterazine-dependent luciferases (isoforms MLuc 164 and MLuc39) from the marine copepod Metridia longa were expressed as inclusion bodies in E. coli cells, dissolved in 6 M guanidinium chloride and folded in conditions developed for proteins containing intramolecular disulfide bonds. One of them (MLuc09) was obtained in an active monomeric form with a high yield. The luciferase bioluminescence is found to be initiated not only by free coelenterazine, but also by Ca2+-dependent coelenterazine-binding protein (CBP) of Renilla muelleri on Ca2+ addition. The use of CBP as a "substrate" provides higher light emission and simultaneously the lower level of background. The high purity MLuc39 can be detected down to attomol with a linear range extending over 5 orders of magnitude. The MLuc39 reveals also a high stability towards heating and chemical modification; the chemically synthesized biotinylated derivatives of the luciferase preserve 35-40% of the initial activity The luciferase applicability as an in vitro bioluminescent reporter is demonstrated in model tandem bioluminescent solid-phase microassay combining the Ca2+-regulated photoprotein obelin and the Metridia luciferase.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Semyonov, Denis A., Eltsov, Ilia, V, Nechipurenko, Yury D.
Заглавие : A New Bias Site for Epigenetic Modifications: How Non-Canonical GC Base Pairs Favor Mechanochemical Cleavage of DNA
Колич.характеристики :8 с
Коллективы : program of the Presidium of the Russian Academy of Sciences for Molecular and Cellular Biology; Program of Fundamental Research for State Academies for years 2013-2020 [01201363818]; Russian Ministry of Science and High Education [05.604.21.0234, RFMEFI60419X0234]
Место публикации : Bioessays: WILEY, 2020. - Vol.: 42, Issue: 11. - Ст.2000051. - ISSN 0265-9247, DOI 10.1002/bies.202000051. - ISSN 1521-1878(eISSN)
Примечания : Cited References:55. - The authors thank Dr. Dmitry M. Graifer, Prof. Vladimir A. Reznikov, and Dr. Anastasia Anashkina, for useful discussion. The authors thank the Prof. D. R. Forsdyke whose advice on improving the style of the manuscript was very useful. This work was supported by the program of the Presidium of the Russian Academy of Sciences for Molecular and Cellular Biology and the Program of Fundamental Research for State Academies for years 2013-2020, project no. 01201363818. The work was supported by the Russian Ministry of Science and High Education (grant #05.604.21.0234, unique number RFMEFI60419X0234)
Предметные рубрики: NUCLEIC-ACIDS
CYTOSINE
MUTAGENESIS
N-4-METHOXYCYTOSINE
Аннотация: Properties of non-canonical GC base pairs and their relations with mechanochemical cleavage of DNA are analyzed. A hypothesis of the involvement of the transient GC wobble base pairs both in the mechanisms of the mechanochemical cleavage of DNA and epigenetic mechanisms involving of 5-methylcytosine, is proposed. The hypothesis explains the increase in the frequency of the breaks of the sugar-phosphate backbone of DNA after cytosines, the asymmetric character of these breaks, and an increase in break frequency in CpG after cytosine methylation. As an alternative hypothesis, probable implication of GC(+)Hoogsteen base pairs is considered, which now exemplify the best-studied non-canonical GC base pairs in the DNA double helix. Also see the video abstract here https://youtu.be/EUunVWL0ptw.
WOS,
Файл: доступ к статье - 0
https://onlinelibrary.wiley.com/doi/epdf/10.1002/bies.202000051
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