Isolation of DNA polymerase alpha from germinated wheat embryos.
DNA polymerase alpha from germinated wheat embryos was purified by ammonium sulphate fractionation, chromatography on DEAE-Toyopearl, followed by phosphocellulose and heparin Sepharose columns. The specific activity of the purified enzyme was more than 60,000 units/mg. It belongs to the alpha-type according to the large molecular mass, high sensitivity to NEM, aphidicoline, 200 mM KCl, low sensitivity to ethidium bromide and the absence of inhibition by ddTTP. DNA polymerase alpha consists of four subunits as shown by SDS-PAGE and seems to be homogeneous under non-denaturing conditions.
Balmukhanov
T S
TS
M.A. Ajtkhozhin Institute of Molecular Biology and Biochemistry, Kazakh Academy of Sciences, Alma-Ata, USSR.
Erekenov
A M
AM
Ajtkhozhina
N A
NA
D000645:Ammonium Sulfate; D005591:Chemical Fractionation; D002845:Chromatography; D004257:DNA Polymerase II / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D054306:Dideoxynucleotides; D004591:Electrophoresis, Polyacrylamide Gel; D005033:Ethylmaleimide / Q000494:pharmacology; D008970:Molecular Weight; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D013942:Thymine Nucleotides / Q000494:pharmacology; D014908:Triticum / Q000201:enzymology*
D054306:Dideoxynucleotides; D013942:Thymine Nucleotides; D010664:Phenylmethylsulfonyl Fluoride; D004257:DNA Polymerase II; C017216:2',3'-dideoxythymidine triphosphate; D005033:Ethylmaleimide; D000645:Ammonium Sulfate