2016
DOI: 10.1111/php.12666
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A Novel Streptavidin–luciferase Fusion Protein: Preparation, Properties and Application in Hybridization Analysis of DNA

Abstract: A streptavidin-luciferase fusion protein comprising the thermostable mutant form of firefly luciferase Luciola mingrelica and minimal core streptavidin was constructed. The streptavidin-luciferase fusion was mainly produced in a tetrameric form with high luciferase and biotin-binding activities. It was shown that fusion has the same K values for ATP and luciferin and the bioluminescence spectra as initial luciferase. The linear dependence of the bioluminescence signal on the content of the fusion was observed … Show more

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Cited by 5 publications
(4 citation statements)
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“…The sensitivity of the immunoassay using the heterologous bridge combination system devised in this study indicates that similar methods would be useful as highly sensitive immunoassays for other small peptides. In addition, this assay can be more simplified by using luciferase–streptavidin or obelin–streptavidin fusing proteins published recently . There is a critical need to continue establishment of valid and reliable methods to measure oxytocin in plasma and other biological fluids.…”
Section: Resultsmentioning
confidence: 99%
“…The sensitivity of the immunoassay using the heterologous bridge combination system devised in this study indicates that similar methods would be useful as highly sensitive immunoassays for other small peptides. In addition, this assay can be more simplified by using luciferase–streptavidin or obelin–streptavidin fusing proteins published recently . There is a critical need to continue establishment of valid and reliable methods to measure oxytocin in plasma and other biological fluids.…”
Section: Resultsmentioning
confidence: 99%
“…К ним относятся высокий выход готового препарата, его стоимость, воспроизводимость и технологичность; полученные препараты, как правило, имеют гомогенный состав, известную стехиометрию и сохраняют функциональную активность. Современные подходы биотехнологической науки практически решили проблему получения рекомбинантных ферментов (таких, как пероксидаза хрена (ПХ), щелочная фосфатаза и люцифераза), использующихся в методах ИФА в качестве маркёров, различных антител и всевозможных антигенов белковой природы [15][16][17][18][19][20][21][22]. Ранее для изучения роли аутоантител к β 1 -адренорецептору в развитии ДКМП был успешно использован рекомбинантный химерный белок, состоящий из глутатион-Sтрансферазы и второй внеклеточной петли, соответствующей аминокислотным остаткам 195-225 β 1 -адренорецептора человека [10].…”
Section: Enzyme-linked Immunosorbent Assay Of Autoantibodies Against ...unclassified
“…Получение рекомбинантных конъюгатов -задача сложная, поскольку на сегодняшний день невозможно достоверно предсказать структуру получаемого конъюгата. В результате из-за неправильного фолдинга двух составных частей химерного белка возможна потеря функциональной активности [19][20][21][22] Таким образом, был разработан метод ИФА для определения аутоантител к β 1адренорецептору человека с использованием рекомбинатного химерного антигена, содержащего участок 2-й внеклеточной петли рецептора.…”
Section: результаты и их обсуждениеunclassified
“…Polyacrylamide gel is used quite often [103] despite its toxicity. Other carriers such as PVA-based cryogels [104,105], ENT/ENTP photocrosslinked polymers (polymer mixture based on polyethylene or polypropylene glycol, hydroxyethyl acrylate, and isophorone diisocyanate, which is polymerized under near UV irradiation) or modified PVA [106], granules and films based on polyvinyl chloride [107], hydrogels based on chitosan [108][109][110], polycarbamoyl sulfonate [111,112], and polyurethane [113,114], peptide polymers [115], biotin-avidin linkers [116][117][118], solgel matrices based on aluminum oxide [119] or composite polymers [120][121][122][123], as well as electropolymeriz able polyaniline films [124][125][126][127] and other compounds [128,129], and nanostructured materials (including carbon nanotubes and metal nanoparticles) [130][131][132][133][134][135][136][137] should also be noted. A common disadvantage of encapsulation in gels and polymers is diffusion restrictions imposed by the carrier nature, which in some cases appreciably reduce the rate of biocatalysis and the activity of biocomponents.…”
Section: Encapsulation Of Cells and Microorganismsmentioning
confidence: 99%