DOI: 10.1111/j.1432-1033.1969.tb19624.x. Extraction and Purification of DNA‐Dependent RNA Polymerase from Rat Liver Nuclei

Extraction and Purification of DNA‐Dependent RNA Polymerase from Rat Liver Nuclei

10.1111/j.1432-1033.1969.tb19624.x

Crossref journal-article

Wiley

European Journal of Biochemistry (311)

Abstract

The enzyme RNA polymerase is difficult to extract from most mammalian cells due to its firm association with chromatin. By a combination of intensive ultrasonic treatment and extraction of the nuclei with 0.75 M NaCl at 0° and pH 8.4 for 1.5 hours approximately 80% of the enzyme can be extracted in a soluble, DNA‐dependent form. The extracted enzyme has been purified by DEAE‐cellulose chromatography and centrifugation on sucrose gradients (5–20%, w/v) containing 10% glycerol. A purification of approximately 250‐fold has been achieved in the peak fraction of the gradient. The enzyme requires all four nucleoside triphosphates, mercaptoethanol, a DNA template and either Mg++ or Mn++, the latter being the preferred ion. Enzyme activity can be stimulated by (NH4)2SO4. No direct stimulation of enzyme activity by cortisol phosphate could be observed.

Bibliography

Seifart, K. H., & Sekeris, C. E. (1969). Extraction and Purification of DNAâDependent RNA Polymerase from Rat Liver Nuclei. European Journal of Biochemistry, 7(3), 408â412. Portico.

Authors 2

K. H. Seifart (first)
C. E. Sekeris (additional)

References 18 Referenced 31

10.1073/pnas.46.8.1020
10.1016/0006-291X(65)90128-2
10.1038/203150a0
10.1016/0006-291X(65)90102-6
10.1016/0005-2787(67)90144-X
{'key': 'e_1_2_2_7_2', 'first-page': '1555', 'volume': '348', 'author': 'Seifart K. H.', 'year': '1967', 'journal-title': "Hoppe-Seyler's Z. Physiol. Chem."} / Hoppe-Seyler's Z. Physiol. Chem. by Seifart K. H. (1967)
10.1016/0005-2787(67)90045-7
10.1016/S0021-9258(18)96755-4 / J. Biol. Chem. by Ballard P. L. (1966)
10.1016/S0021-9258(19)52451-6
10.1016/S0021-9258(18)70943-5
10.1016/0005-2787(67)90092-5
10.1016/0006-3002(61)91042-3
10.1016/S0076-6879(62)05183-6
{'key': 'e_1_2_2_15_2', 'first-page': '112', 'volume-title': 'Partition of Cell particles and Macromolecules', 'author': 'Albertsson P.', 'year': '1960'} / Partition of Cell particles and Macromolecules by Albertsson P. (1960)
10.1073/pnas.59.1.262
10.1016/S0022-2836(64)80076-0
10.1515/bchm2.1967.348.1.902
10.1515/bchm2.1968.349.2.1126 / Hoppe-Seyler's Z. Physiol. Chem. by Neuhoff V. (1968)

Dates

Type	When
Created	20 years, 5 months ago (March 3, 2005, 10:16 a.m.)
Deposited	1 year, 9 months ago (Nov. 23, 2023, 2:53 a.m.)
Indexed	1 year, 9 months ago (Nov. 23, 2023, 3:10 a.m.)
Issued	56 years, 7 months ago (Jan. 1, 1969)
Published	56 years, 7 months ago (Jan. 1, 1969)
Published Online	20 years, 5 months ago (March 3, 2005)
Published Print	56 years, 7 months ago (Jan. 1, 1969)

Funders 0

None

BibTeX

@article{Seifart_1969, title={Extraction and Purification of DNA‐Dependent RNA Polymerase from Rat Liver Nuclei}, volume={7}, ISSN={1432-1033}, url={http://dx.doi.org/10.1111/j.1432-1033.1969.tb19624.x}, DOI={10.1111/j.1432-1033.1969.tb19624.x}, number={3}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={Seifart, K. H. and Sekeris, C. E.}, year={1969}, month=jan, pages={408–412} }

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