[DNA-polymerase from sea urchin (Strongylocentrotus intermedius. Embryos].
Using DEAE-cellulose chromatography, three peaks of the DNA-polymerase activity were found in a homogenate of the sea urchin Strongylocentrotus intermedius embryos at stage 32 of the blastomer. The isolation and purification of DNA-polymerase making up the bulk of he DNA-synthesizing activity of the sea urchin embryo cells included fractionation by ammonium sulfate, chromatography on DEAE-cellulose, hydroxyapatite and DNA-cellulose, resulting in a 2750-fold purification of the enzyme. The enzyme activity requires the presence of two-chain DNA activated by pancreatic DNAse, four dNTP and Mg2+. The enzyme is inhibited by a high ionic strength (150 mM KCl or NaCL) and the sulfhydryl reagent--N-ethylmaleimide; the pH optimum is 8.0. The molecular weight of the enzyme as determined by gel-filtration is about 150 000. It is assumed that the enzyme under study can be related to DNA-polymerases of the alpha-type.
D000818:Animals; D004257:DNA Polymerase II / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000201:enzymology; D005260:Female; D007700:Kinetics; D008970:Molecular Weight; D012617:Sea Urchins / Q000201:enzymology*
D004257:DNA Polymerase II; D004259:DNA-Directed DNA Polymerase
Dnk-polimeraza iz embrionov morskogo ezha Strongylocentrotus intermedius. Ochistka i nekotorye svoĭstva.