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Renaturation, Activation, and Practical Use of Recombinant Duplex-Specific Nuclease from Kamchatka Crab


V. E. Anisimova, D. V. Rebrikov, P. A. Zhulidov, D. B. Staroverov, S. A. Lukyanov, and A. S. Shcheglov*

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117997 Moscow, Russia; fax: (7-495) 330-7056; E-mail: jukart@mail.ru

* To whom correspondence should be addressed.

Received November 2, 2005; Revision received January 26, 2006
We overexpressed duplex-specific nuclease (DSN) from Kamchatka crab in Escherichia coli cells and developed procedures for purification, renaturation, and activation of this protein. We demonstrated identity of the properties of the native and recombinant DSN. We also successfully applied the recombinant DSN for full-length cDNA library normalization.
KEY WORDS: duplex-specific nuclease (DSN), sugar nonspecific nuclease, renaturation, activation, specificity

DOI: 10.1134/S0006297906050075