Abstract
Salt tolerance is an important property of duplex-specific nuclease (DSN). DSN with high salt tolerance can be more widely used in genetic engineering, especially in the production of nucleic acid drugs. To improve the salt tolerance of DSN, we selected five DNA-binding domains from extremophilic organisms, which have been shown the ability to improve salt tolerance of DNA polymerases and nucleases. The experimental results demonstrated that the fusion protein TK-DSN produced by fusing a N-terminal DNA-binding domain, which comprised two HhH (helix–hairpin–helix) motifs domain from an extremely halotolerant bacterium Thioalkalivibrio sp. K90mix, has a significantly improved salt tolerance. TK-DSN can tolerate the concentration of NaCl up to 800 mM; in addition, the ability of digesting DNA was also enhanced during in vitro transcription and RNA purification. This strategy provides the method for the personalized customization of biological tool enzymes for different applications.
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The data that support the findings of this study are available from the corresponding author by reasonable request.
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Acknowledgements
The authors gratefully acknowledge Dr. Henghao Xu, Dr. Zhidan Luo for their comments on the manuscript.
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This work was supported by the Priority Academic Program Development of Jiangsu Higher Education Institutions of China.
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Hu, W., Wang, J., Li, J. et al. Improvement of duplex-specific nuclease salt tolerance by fusing DNA-binding domain of DNase from an extremely halotolerant bacterium Thioalkalivibrio sp. K90mix. Extremophiles 27, 11 (2023). https://doi.org/10.1007/s00792-023-01296-1
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DOI: https://doi.org/10.1007/s00792-023-01296-1