scholarly journals DENSITY GRADIENT ISOLATION OF RAT LIVER NUCLEI WITH HIGH DNA CONTENT

1963 ◽  
Vol 17 (2) ◽  
pp. 231-236 ◽  
Author(s):  
Richard F. Fisher ◽  
David J. Holbrook ◽  
J. Logan Irvin
Keyword(s):  
Density Gradient ◽  
Rat Liver ◽  
Dna Content ◽  
Phosphorus Content ◽  
Sucrose Solution ◽  
Normal Liver ◽  
Sucrose Density Gradient ◽  
Pellet Fraction ◽  
Rat Liver Nuclei ◽  
Liver Nuclei

Rat liver nuclei, after preliminary isolation in 2.2 molar sucrose solution, were separated into density classes by centrifugation at 95,000 g for 45 to 85 minutes in a sucrose density gradient (density range, 1.28 to 1.33). Nuclei from normal liver separated into three bands with average DNA phosphorus content per nucleus of 0.67, 0.84, and 0.93 picogram for top, middle, and bottom bands, respectively. Nuclei from regenerating liver (26 hours after one-third hepatectomy) yielded three bands and a pellet fraction with average DNA phosphorus content per nucleus of 0.76, 1.02, 1.38, and 1.51 picograms (top to bottom of tube). This method appears capable of yielding nuclei which have increased their DNA content prior to mitosis, and this procedure should be valuable in studies of biochemical changes which occur in nuclei preparing for mitosis.

Labelling of Proteins by Isolated Rat Liver Nuclei

1972 ◽  
Vol 50 (2) ◽  
pp. 190-199 ◽  
Author(s):  
K. M. Anderson ◽  
M. Slavik ◽  
O. P. Elebute
Keyword(s):  
Protein Synthesis ◽  
Rat Liver ◽  
Nuclear Protein ◽  
Sucrose Solution ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Rat Liver Nuclei ◽  
Teleological Argument ◽  
Liver Nuclei ◽  
Triton X

Rat liver nuclei, isolated in hypertonic sucrose solution and washed with Triton X-100, incorporate radioactive amino acids into hot trichloroacetic acid insoluble materials.Optical and biochemical evidence of nuclear purity is presented. The temperature-dependent incorporation continued for 20–30 min, and was proportional to the concentrations of both nuclear protein between 0.5–1.5 mg/ml, and radioactive amino acid. The radioactive product was degraded by pronase, and a number of inhibitors reduced incorporation, but only if present at [Formula: see text]. Proteins extracted from labelled nuclei and microsomes and examined by sodium dodecyl sulfate–polyacrylamide gel electrophoresis at pH 7.2 exhibited different patterns of radioactivity. This provides further support for the concept of protein synthesis intrinsic to rat liver nuclei.A teleological argument for the function of nuclear protein synthesis is discussed.

Age And Polyploidy Of Rat Liver NUCLEI As Measured By Volume And DNA Content

1959 ◽  
Vol 14 (1) ◽  
pp. 2-8 ◽  
Author(s):  
J. A. Falzone ◽  
C. H. Barrows ◽  
N. W. Shock
Keyword(s):  
Rat Liver ◽  
Dna Content ◽  
Rat Liver Nuclei ◽  
Liver Nuclei

Characterization and Partial Purification of Heterodisperse Polysomal RNAs in Normal and Neoplastic Cells

1972 ◽  
Vol 58 (2) ◽  
pp. 71-94
Author(s):  
Ada Sacchi ◽  
Gianni Chinali ◽  
Susetta Pons ◽  
Michela Galdieri ◽  
Piero Cammarano
Keyword(s):  
Size Distribution ◽  
Density Gradient ◽  
Rat Liver ◽  
Sucrose Density Gradient ◽  
Partial Purification ◽  
Messenger Rnas ◽  
Alkaline Ph ◽  
Sedimentation Profile ◽  
Molecular Weights ◽  
Phenol Extraction

The size distribution of cytoplasmic messenger RNAs (m-RNA) has been studied in rat liver and in monodifferentiated cells (mouse reticulocytes and myelomas). It has been found that the RNA which exhibits a « rapid turnover » and a polydisperse profile of radioactivity is refractory to phenol extraction. This property has been exploited to selectively isolate m–RNA from the phenol residue by means of an extraction at an alkaline pH. The sucrose density gradient profiles of m–RNA isolated from monodifferentiated cells show monodisperse peaks having the sedimentation coefficients expected on the basis of the molecular weights of monocistronic messages for α and β chains of hemoglobin (reticulocytes) and L and H chains of immunoglobulin (myelomas). The sedimentation profile of cytoplasmic m–RNA associated with rat liver polysomes shows a much broader distribution, with sedimentation coefficients ranging from 8 S to 28 S.

Sign in / Sign up

Export Citation Format

Share Document