Thermolabile duplex-specific nuclease |
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Authors: | Veronika E. Anisimova Ekaterina V. Barsova Ekaterina A. Bogdanova Sergey A. Lukyanov Alex S. Shcheglov |
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Affiliation: | (1) Shemiakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, 117871 Moscow, Russia;(2) Evrogen JSC, Miklukho-Maklaya 16/10, 117871 Moscow, Russia |
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Abstract: | Using random mutagenesis of the gene encoding duplex-specific nuclease from the king crab we found a new mutant that retained all properties of the wild-type protein, but exhibited a much lower thermal stability. This enzyme, denoted thermolabile duplex-specific nuclease (DSN-TL), exhibits high processivity and selective cleavage of dsDNA. The inactivation temperature for DSN-TL is 15–20°C lower than that of the widely used DNase I and shrimp nuclease, and its catalytic activity is more than 10 times higher. Moreover, DSN-TL is resistant to proteinase K treatment. These properties make DSN-TL very useful for removing genomic DNA from RNA samples intended for quantitative RT-PCR. |
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Keywords: | Bovine DNase I Duplex-specific nuclease Shrimp nuclease Thermolabile |
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