Renaturation, activation, and practical use of recombinant duplex-specific nuclease from Kamchatka crab.
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.
Anisimova
V E
VE
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
Rebrikov
D V
DV
Zhulidov
P A
PA
Staroverov
D B
DB
Lukyanov
S A
SA
Shcheglov
A S
AS
D000818:Animals; D001483:Base Sequence; D003386:Brachyura / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics; D004720:Endonucleases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation; D004795:Enzyme Stability; D009692:Nucleic Acid Heteroduplexes / Q000378:metabolism; D017510:Protein Folding; D020673:Protein Renaturation; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D018076:DNA, Complementary; D009692:Nucleic Acid Heteroduplexes; D011994:Recombinant Proteins; D004720:Endonucleases