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; A S Shcheglov

doi:10.1134/s0006297906050075

Renaturation, activation, and practical use of recombinant duplex-specific nuclease from Kamchatka crab

Biochemistry (Mosc). 2006 May;71(5):513-9. doi: 10.1134/s0006297906050075.

Authors

V E Anisimova¹, D V Rebrikov, P A Zhulidov, D B Staroverov, S A Lukyanov, A S Shcheglov

Affiliation

¹ Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.

PMID: 16732729
DOI: 10.1134/s0006297906050075

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Base Sequence
Brachyura / enzymology*
Brachyura / genetics
Brachyura / metabolism
Cloning, Molecular / methods
DNA, Complementary / genetics
Endonucleases / chemistry
Endonucleases / genetics
Endonucleases / metabolism*
Enzyme Activation
Enzyme Stability
Nucleic Acid Heteroduplexes / metabolism
Protein Folding
Protein Renaturation
Recombinant Proteins / genetics
Recombinant Proteins / isolation & purification
Recombinant Proteins / metabolism*

Substances

DNA, Complementary
Nucleic Acid Heteroduplexes
Recombinant Proteins
Endonucleases