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Thermolabile duplex-specific nuclease

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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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Acknowledgments

We are very grateful to Prof. NB Gusev for valuable advice during the preparation of this manuscript. This work was supported by the Evrogen JSC (Russia) and Presidium RAS (grant No. 34-405/06-60).

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Authors and Affiliations

  1. Shemiakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, 117871, Moscow, Russia

    Veronika E. Anisimova, Ekaterina V. Barsova, Ekaterina A. Bogdanova, Sergey A. Lukyanov & Alex S. Shcheglov

  2. Evrogen JSC, Miklukho-Maklaya 16/10, 117871, Moscow, Russia

    Veronika E. Anisimova

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  1. Veronika E. Anisimova
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  2. Ekaterina V. Barsova
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  3. Ekaterina A. Bogdanova
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  4. Sergey A. Lukyanov
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  5. Alex S. Shcheglov
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Correspondence to Veronika E. Anisimova.

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Anisimova, V.E., Barsova, E.V., Bogdanova, E.A. et al. Thermolabile duplex-specific nuclease. Biotechnol Lett 31, 251–257 (2009). https://doi.org/10.1007/s10529-008-9850-y

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  • DOI: https://doi.org/10.1007/s10529-008-9850-y

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