title: Thermolabile duplex-specific nuclease.
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 degrees 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.
authors.affiliation: Shemiakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, 117871, Moscow, Russia. evanika@yandex.ru
authors.forename: Veronika E
authors.initials: VE
authors.lastname: Anisimova
authors.forename: Ekaterina V
authors.initials: EV
authors.lastname: Barsova
authors.forename: Ekaterina A
authors.initials: EA
authors.lastname: Bogdanova
authors.forename: Sergey A
authors.initials: SA
authors.lastname: Lukyanov
authors.forename: Alex S
authors.initials: AS
authors.lastname: Shcheglov
mesh_terms: D000818:Animals; D001665:Binding Sites; D003386:Brachyura / Q000201:enzymology* / Q000235:genetics; D002384:Catalysis; D004247:DNA / Q000737:chemistry* / Q000235:genetics*; D003851:Deoxyribonucleases / Q000737:chemistry* / Q000235:genetics*; D004789:Enzyme Activation; D004795:Enzyme Stability; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods; D013329:Structure-Activity Relationship; D013696:Temperature
chemical_list: D004247:DNA; D003851:Deoxyribonucleases