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Violet and greenish photoprotein obelin mutants for reporter applications in dual-color assay [Text] / L. A. Frank [et al.] // Anal. Bioanal. Chem. - 2008. - Vol. 391, Is. 8. - P2891-2896, DOI 10.1007/s00216-008-2223-5. - Cited References: 22 . - ISSN 1618-2642

РУБ Biochemical Research Methods + Chemistry, Analytical
Рубрики:
ANGSTROM RESOLUTION
   RECOMBINANT OBELIN
   CRYSTAL-STRUCTURE
   BIOLUMINESCENCE
   AEQUORIN
   IMMUNOASSAY
   EXPRESSION
   CDNA
   PURIFICATION
   CLONING
Кл.слова (ненормированные):
Ca(2+)-regulated photoprotein -- bioluminescence -- dual-color assay
Аннотация: Two kinds of Ca(2+)-regulated photoprotein obelin with altered color of bioluminescence were obtained by active-center amino acid substitution. The mutant W92F-H22E emits violet light (lambda(max)=390 nm) and the mutant Y139F emits greenish light (lambda (max)=498 nm), with small spectral overlap, both display high activity and stability and thus may be used as reporters. For demonstration, the mutants were applied in dual-color simultaneous immunoassay of two gonadotropic hormones-follicle-stimulating hormone and luteinizing hormone. Bioluminescence of the reporters was simultaneously triggered by single injection of Ca(2+) solution, divided using band-pass optical filters and measured with a two-channel photometer. The sensitivity of simultaneous bioluminescence assay was close to that of a separate radioimmunoassay.

Держатели документа:
[Frank, Ludmila A.] Siberian Fed Univ, Krasnoyarsk 660041, Russia
[Frank, Ludmila A.
Borisova, Vasilisa V.
Markova, Svetlana V.
Malikova, Natalia P.
Stepanyuk, Galina A.
Vysotski, Eugene S.] Russian Acad Sci, Inst Biophys, Siberian Branch, Krasnoyarsk 660036, Russia
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Frank, L.A.; Borisova, V.V.; Markova, S.V.; Malikova, N.P.; Stepanyuk, G.A.; Vysotski, E.S.

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Testing soil-like substrate for growing plants in bioregenerative life support systems / J. B. Gros [et al.] // Advances in Space Research. - 2005. - Vol. 36, Is. 7. - P1312-1318, DOI 10.1016/j.asr.2005.05.079 . - ISSN 0273-1177
Кл.слова (ненормированные):
Life support system -- Matter recycling -- Plants -- Soil-like substrate -- Biodiversity -- Correlation methods -- Growth kinetics -- Hormones -- Plants (botany) -- Recycling -- Soils -- Bioregeneration -- Life support system -- Matter recycling -- Soil-like substrate -- Space research
Аннотация: We studied soil-like substrate (SLS) as a potential candidate for plant cultivation in bioregenerative life support systems (BLSS). The SLS was obtained by successive conversion of wheat straw by oyster mushrooms and worms. Mature SLS contained 9.5% humic acids and 4.9% fulvic acids. First, it was shown that wheat, bean and cucumber yields as well as radish yields when cultivated on mature SLS were comparable to yields obtained on a neutral substrate (expanded clay aggregate) under hydroponics. Second, the possibility of increasing wheat and radish yields on the SLS was assessed at three levels of light intensity: 690, 920 and 1150 ?mol m-2 s-1 of photosynthetically active radiation (PAR). The highest wheat yield was obtained at 920 ?mol m-2 s-1, while radish yield increased steadily with increasing light intensity. Third, long-term SLS fertility was tested in a BLSS model with mineral and organic matter recycling. Eight cycles of wheat and 13 cycles of radish cultivation were carried out on the SLS in the experimental system. Correlation coefficients between SLS nitrogen content and total wheat biomass and grain yield were 0.92 and 0.97, respectively, and correlation coefficients between nitrogen content and total radish biomass and edible root yield were 0.88 and 0.87, respectively. Changes in hormone content (auxins, gibberellins, cytokinins and abscisic acid) in the SLS during matter recycling did not reduce plant productivity. Quantitative and species compositions of the SLS and irrigation water microflora were also investigated. Microbial community analysis of the SLS showed bacteria from Bacillus, Pseudomonas, Proteus, Nocardia, Mycobacterium, Arthrobacter and Enterobacter genera, and fungi from Trichoderma, Penicillium, Fusarium, Aspergillus, Mucor, Botrytis, and Cladosporium genera. В© 2005 COSPAR. Published by Elsevier Ltd. All rights reserved.

Scopus
Держатели документа:
LGCB, Universite B. Pascal, CUST, BP206, 63174 Aubiere cedex, France
Environmental Control and Life Support Section, ESA-Estec, Postbus 299, 2200 AG, Noordwijk, Netherlands
Institute of Biophysics (Russian Academy of Sciences, Siberian Branch), Academgorodok, Krasnoyarsk 660036, Russian Federation
Department of Plant Physiology and Biotechnology, Tomsk State University, Lenin av. 36, Tomsk 634050, Russian Federation : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Gros, J.B.; Lasseur, Ch.; Tikhomirov, A.A.; Manukovsky, N.S.; Kovalev, V.S.; Ushakova, S.A.; Zolotukhin, I.G.; Tirranen, L.S.; Karnachuk, R.A.; Dorofeev, V.Yu.

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Bioluminescent immunoassay of thyrotropin and thyroxine using obelin as a label [Text] / L. A. Frank, A. I. Petunin, E. S. Vysotski // Anal. Biochem. - 2004. - Vol. 325, Is. 2. - P240-246, DOI 10.1016/j.ab.2003.11.003. - Cited References: 16 . - ISSN 0003-2697

РУБ Biochemical Research Methods + Biochemistry & Molecular Biology + Chemistry, Analytical
Рубрики:
AEQUORIN
   PURIFICATION
   PROTEIN
   CDNA
Кл.слова (ненормированные):
obelin -- human thyrotropin -- thyroxine -- immunoassay -- bioluminescence
Аннотация: Solid-phase bioluminescent immunoassay of thyroid hormones, human thyrotropin (hTSH) and two forms of thyroxine (T4), whose determinations are vitally important for diagnostics of thyroid diseases and the efficiency of treatment, is described. The recombinant obelin, a Ca2+-regulated photoprotein originally derived from the luminous marine hydroid Obelia longissima, is employed as a bioluminescent label. To produce obelin conjugates with anti-hTSH, anti-T4 immunoglobulins (IgG), and T4, additional SH groups are introduced into the obelin molecule using Traut's reagent (2-iminothiolane) and then obelin possessing extra SH groups is conjugated with succinimidyl 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate-activated IgGs or T4. The total yield of obelin conjugates determined by luminescent activity is 60-65% after all chemical and purification procedures. The obtained conjugates are stable to lyophilization and in solution for at least 9 months at 4 degreesC, with loss of activity not exceeding 10%. The application of obelin conjugates for determination of the hTSH, total T4, and free T4 in standard, control, and patient sera displays high sensitivity and reproducibility of results. The results of bioluminescent immunoassays are closely comparable to those obtained by the radioimmunoassay method (R = 0.95-0.99). (C) 2003 Elsevier Inc. All rights reserved.

Держатели документа:
Russian Acad Sci, Inst Biophys, Siberian Branch, Krasnoyarsk 660036, Russia
DIAS Ltd, Krasnoyarsk 660036, Russia
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Frank, L.A.; Petunin, A.I.; Vysotski, E.S.

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