Тип публикации: статья из журнала

Год издания: 1995

Аннотация: Obelin mRNA obtained in vitro with the aid of SP6 RNA polymerase was translated in a wheat germ cell-free system. Only the polypeptide with a molecular mass of about 20 kDa was synthesized. The activation of apoobelin with a synthetic coelenterazine revealed a luminescence activity initiated by calcium. The specific activity was 3.Показать полностью6 +/- 0.4 x 10(15)photons per mg of the in vitro synthesized obelin (k=6.9s(-1)). The luminescence of the obelin was in a good correlation with the protein concentration calculated by the incorporation of [14C]Leu. The determination of the amount of de novo synthesized obelin based on measurement of its luminescence is one-thousand times more sensitive than the approach based on the incorporation of labeled amino acid. Thus, obelin mRNA has some advantages for evaluating the efficiency of cell-free translation when compared with standard methods.

Журнал: Analytical Biochemistry

Выпуск журнала: Т. 231, № 1

Номера страниц: 34-39

ISSN журнала: 00032697

Издатель: Academic Press

• Matveev S.K. (Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences)
• Illarionov B.A. (Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences)
• Vysotsky E.S. (Institute of Biophysics, Siberian Branch of the Russian Academy of Sciences)
• Bondar V.S. (Institute of Biophysics, Siberian Branch of the Russian Academy of Sciences)
• Markova S.V. (Institute of Biophysics, Siberian Branch of the Russian Academy of Sciences)
• Alakhov Y.B. (Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences)

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