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Overexpression and refolding of thioredoxin/TRAIL fusion from inclusion bodies and further purification of TRAIL after cleavage by enteropeptidaseстатья
Информация о цитировании статьи получена из
Web of Science,
Scopus
Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 18 июля 2013 г.
- Авторы: Gasparian M.E., Ostapchenko V.G., Yagolovich A.V., Tsygannik I.N., Chernyak B.V., Dolgikh D.A., Kirpichnikov M.P.
- Журнал: Biotechnology Letters
- Том: 29
- Номер: 10
- Год издания: 2007
- Издательство: Springer Nature
- Местоположение издательства: Switzerland
- Первая страница: 1567
- Последняя страница: 1573
- DOI: 10.1007/s10529-007-9446-y
- Аннотация: The human TRAIL gene (encoding residues 114-281) was synthesized by PCR and cloned into plasmid pET-32a. High level expression (1.5 g l(-1)) of thioredoxin/TRAIL fusion was achieved in Escherichia coli strain BL21(DE3), mainly as inclusion bodies. Refolded fusion thioredoxin/TRAIL was cleaved by enteropeptidase and TRAIL was separated from thioredoxin on Ni-NTA agarose. High yield (400 mg l(-1)) of TRAIL without N-terminal methionine and His tag was obtained. Sedimentation coefficient demonstrated that 98% of TRAIL formed trimers. TRAIL formed crystals of space group P3 (1) with unit-cell dimensions a = b = 72.5 A, c = 141.5 A. Apoptosis induced in HeLa cells by purified TRAIL was 5-fold enhanced by emetine.
- Добавил в систему: Черняк Борис Викторович