STUDIES ON THE RELATIONSHIP BETWEEN VIRUS AND HOST CELL: THE PREPARATION OF T2r+ BACTERIOPHAGE LABELLED WITH RADIOACTIVE PHOSPHORUS

1950 ◽  
Vol 28e (6) ◽  
pp. 281-288 ◽  
Author(s):  
S. M. Lesley ◽  
R. C. French ◽  
A. F. Graham
Keyword(s):  
Nucleic Acid ◽  
Host Cell ◽  
Inorganic Phosphate ◽  
Specific Activity ◽  
High Specific Activity ◽  
Inorganic Phosphorus ◽  
Radioactive Phosphorus ◽  
Specific Activities ◽  
The Relationship ◽  
Escherichia Coli B

T2r+ bacteriophage grown in its host, Escherichia coli B, in broth medium in the presence of radioactive inorganic phosphorus was labelled with the isotope. Purified suspensions of this virus had specific activities up to 50,000 c.p.m. per μgm. P. There was little or no exchange of P32 between virus and inorganic phosphate. Chemical analysis showed that at least 98% of the virus phosphorus was contained in nucleic acid; of the nucleic acid phosphorus 95.5% was associated with desoxypentose nucleic acid and 4.5% with pentose nucleic acid. More than 99% of the radioactivity of the labelled bacteriophage was contained in the nucleic acid fraction. Preparations of bacteriophage were obtained with sufficiently high specific activity to enable metabolism experiments to be carried out on the growth of the labelled virus in the host cell.

scholarly journals STUDIES OF PHOSPHORUS METABOLISM IN MAN: II. A STUDY OF THE PERMEABILITY OF THE HUMAN ERYTHROCYTE TO INORGANIC PHOSPHATE IN VITRO BY THE USE OF RADIOACTIVE PHOSPHATE (P32)

Blood ◽  
1948 ◽  
Vol 3 (12) ◽  
pp. 1472-1477 ◽  
Author(s):  
F. H. L. TAYLOR ◽  
S. M. LEVENSON ◽  
M. A. ADAMS ◽  
MARY KENDRICK
Keyword(s):  
Human Erythrocyte ◽  
Inorganic Phosphate ◽  
Sodium Phosphate ◽  
Specific Activity ◽  
Soluble Fraction ◽  
High Specific Activity ◽  
Phosphorus Metabolism ◽  
Phosphate Exchange ◽  
Inorganic Fraction

Abstract 1. Phosphate exchange in red cells and plasma was studied in vitro using P32 in the form of sodium phosphate as a tracer. 2. No phosphate was added other than the isotopic preparation which was of high specific activity. 3. Inorganic phosphate exchanged freely between the plasma and the erythrocytes at 37.5 C. in a period of four hours. Minimal transfer occurred at 7 C. 4. Most of the added P32 which passed into the erythrocytes during this time remained in the inorganic fraction, less than 15 per cent being found in the organic acid soluble fraction. 5. The specific activity of the inorganic phosphate of the erythrocytes was equal to or greater than that obtaining for the inorganic phosphate of the plasma at the end of the four hour incubation period at 37.5 C.

THE RELATIONSHIP BETWEEN CORTICOSTERONE SYNTHESIS FROM ENDOGENOUS PRECURSORS AND FROM ADDED RADIOACTIVE PRECURSORS BY RAT ADRENAL TISSUE IN VITRO

1966 ◽  
Vol 36 (3) ◽  
pp. 231-238 ◽  
Author(s):  
G. P. VINSON
Keyword(s):  
Specific Activity ◽  
Single Point ◽  
Biosynthetic Pathways ◽  
Activity Change ◽  
Adrenal Tissue ◽  
Specific Activities ◽  
The Relationship ◽  
Synthesis Of Hormones ◽  
Venous Plasma

SUMMARY A kinetic study of corticosterone recovered during incubation of rat adrenal tissue with [4-14C]progesterone and [16-3H]pregnenolone shows that both its 3H: 14C ratio and its specific activity change with time throughout incubation. Corticosterone is itself metabolized, and hypotheses based on the rate of production of corticosterone may be therefore deceptive when the amount is measured at a single point in time. Thus disparities between the specific activities of products and intermediates may be expected which alone do not necessarily indicate differences between the biosynthetic pathways involved in production from endogenous precursors and those from added radioactive precursors. Other experiments suggest that steroids in incubation media, in concentrations comparable with those found in adrenal venous plasma, do not inhibit the continued synthesis of hormones.

NON-LETHAL ASSIMILATION AND DISTRIBUTION OF RADIOACTIVE PHOSPHORUS IN SERRATIA MARCESCENS

1964 ◽  
Vol 10 (3) ◽  
pp. 317-322 ◽  
Author(s):  
G. E. Myers ◽  
R. G. L. McCready
Keyword(s):  
Serratia Marcescens ◽  
Culture Medium ◽  
Specific Activity ◽  
Test Organism ◽  
High Specific Activity ◽  
Lethal Effect ◽  
Optimal Conditions ◽  
Radioactive Phosphorus ◽  
Lipid Fractions ◽  
Cell Fractions

Optimal conditions for non-lethal assimilation of radioactive phosphorus by Serratia marcescens have been studied. Ten microcuries or less of P32per 300 ml of medium has no significant lethal effect on the test organism. The presence of 1% w/v glucose in the culture medium stimulates phosphorus assimilation causing an increase of approximately 34%. Phosphorus assimilation over a 24 hour period increases by approximately 50% when cultures are incubated at 30 °C rather than 37 °C.The concentration of P32in the various cellular constituents has been determined at various intervals during 24 hour incubation. Incorporation of P32in the acid-soluble and lipid fractions of the cell begins almost immediately whereas there is a slight delay prior to incorporation into the R.N.A. and longer delay before incorporation into D.N.A. Sixty to 65% of the P32assimilated by the bacterial cell is incorporated into the nucleic acids. Although R.N.A. has the highest specific activity of the cell fractions studied, the phospholipid fraction also has high specific activity.

scholarly journals Oestrogen Sulfotransferase: Isolation of a High Specific Activity Species from Bovine Placenta

1988 ◽  
Vol 41 (3) ◽  
pp. 333 ◽  
Author(s):  
Stephen S Moore ◽  
EOP Thompson ◽  
Allan R Nash
Keyword(s):  
Mammary Cancer ◽  
Specific Activity ◽  
High Specific Activity ◽  
Gel Permeation Chromatography ◽  
Salt Precipitation ◽  
Bovine Placenta ◽  
Steroid Binding Proteins ◽  
Specific Activities ◽  
Steroid Binding ◽  
Control Of Expression

During the course of a study of the control of expression of steroid-binding proteins in human mammary cancer oestrogen sulfotransferase was isolated from bovine placenta. By a combination of salt precipitation and ion-exchange and gel-permeation chromatography two forms of the enzyme were isolated. The forms, which apparently differ only in charge, have specific activities 100-300 times greater than has previously been reported for the enzyme. Partial peptide sequences of these enzymes are presented.

The effects of diets deficient in phosphorus, phosphorus and vitamin D, or calcium, on the skeleton and teeth of the growing sheep IV. The uptake of radioactive phosphorus by the bones and teeth

1965 ◽  
Vol 65 (1) ◽  
pp. 21-27 ◽  
Author(s):  
M. R. McRoberts ◽  
R. Hill ◽  
A. C. Dalgarno
Keyword(s):  
Vitamin D ◽  
Inverse Relationship ◽  
Specific Activity ◽  
Radioactive Phosphorus ◽  
Specific Activities ◽  
Strong Inverse Relationship

1. Radioactive phosphorus was injected into young sheep that had been fed diets low in either phosphorus, phosphorus and vitamin D or calcium. Forty-eight hours after the injection the animals were killed and samples of bone and teeth were removed for specific activity determinations.2. The specific activities of bone varied according to the site from which it was taken, but in general a strong inverse relationship was found between specific activity and the quality of the skeleton.3. In teeth that were formed during the period the diets deficient in phosphorus or phosphorus and vitamin D were fed, the specific activities of the dentine and enamel were greater than for corresponding tissues from the control animals.

THE UPTAKE OF RADIOACTIVE PHOSPHORUS BY INFLUENZA VIRUS A (PR8 STRAIN)

1950 ◽  
Vol 28e (3) ◽  
pp. 121-134 ◽  
Author(s):  
A. F. Graham ◽  
Laurella McClelland
Keyword(s):  
Influenza Virus ◽  
Specific Activity ◽  
Inorganic Phosphorus ◽  
Experimental Conditions ◽  
Virus Growth ◽  
Radioactive Phosphorus ◽  
Influenza Virus A ◽  
Direct Exchange ◽  
Subsequent Period

Radioactive inorganic phosphorus placed in the allantoic sacs of embryonated eggs three hours after inoculation with influenza virus was incorporated into the structure of the virus during its growth. There was little or no direct exchange between the virus and radioactive inorganic phosphorus. The specific activity of purified labelled virus rose linearly with increasing amounts of radioactive phosphorus administered to the eggs. When radioactive phosphorus was placed in the allantoic sac 48 hr. before inoculation with influenza virus the newly formed labelled virus had a specific activity about 20% higher than when isotope was administered at the same time as virus. As the amount of isotope injected into each infected egg was increased up to 775 μrd. an increasing number of embryos died during the subsequent period of virus growth. The yield of virus from the surviving eggs was not less than from eggs which had not received radioactive phosphorus. Under the experimental conditions described the amount of isotope which could be introduced into influenza virus was not sufficient to permit the use of the marked virus in metabolism experiments in animals or embryonated eggs.

Eine Methode zur Darstellung Tritium-markierter cancerogener polycyclischer Kohlenwasserstoffe mit extrem hoher spezifischer Aktivität

1966 ◽  
Vol 21 (9) ◽  
pp. 855-858 ◽  
Author(s):  
Hans Emmerich ◽  
Peter Schmialek
Keyword(s):  
Polycyclic Aromatic Hydrocarbons ◽  
Thin Layer ◽  
Paper Chromatography ◽  
Aromatic Hydrocarbons ◽  
Radiochemical Purity ◽  
Specific Activity ◽  
High Specific Activity ◽  
Polycyclic Aromatic ◽  
Specific Activities

A method is described for the preparation of tritiated polycyclic aromatic hydrocarbons with extremely high specific activity in the presence of AlCl3 and HTO. The hydrocarbons are purified by preparative thin layer and paper chromatography. The radiochemical purity of the compounds was proved by paper chromatography and dilution analysis. The following specific activities are reached: Anthracene: 54 Curies/mMol, 7,12-Dimethylbenzanthracene: 19 Curies/mMol, 3,4-Benzopyrene: 24 Curies/mMol, and 3-Methylcholanthrene: 44 Curies/mMol.

THE FRACTIONATION OF PHOSPHORUS CONTAINING CONSTITUENTS IN THE ALLANTOIC MEMBRANE OF THE EMBRYONATED EGG

1950 ◽  
Vol 28e (6) ◽  
pp. 271-280 ◽  
Author(s):  
A. F. Graham
Keyword(s):  
Influenza Virus ◽  
Nucleic Acid ◽  
Nucleic Acids ◽  
Total Phosphorus ◽  
Trichloroacetic Acid ◽  
Inorganic Phosphorus ◽  
Radioactive Phosphorus ◽  
Significant Difference ◽  
Phosphorus Containing ◽  
Soluble Phosphorus

A procedure is described which permitted the phosphorus containing constituents in allantoic membranes of embryonated eggs to be separated into fractions as follows: alcohol soluble phosphorus from which the phospholipids were separated, phosphorus soluble in 5% trichloroacetic acid in which inorganic phosphorus was determined, and nucleic acid phosphorus which was further separated into pentose and desoxypentose nucleic acids. This procedure was applied to membranes between 9 and 13 days of age. It was found that the total phosphorus amounted to 9 to 10 mgm. per gm. of dried tissue. Alcohol soluble phosphorus accounted for approximately 28%, acid soluble phosphorus for 37%, and nucleic acid phosphorus for 35% of the total phosphorus. About half the acid soluble phosphorus was inorganic and about 85% of the alcohol soluble phosphorus was associated with phospholipids. These proportions remained essentially constant over the period studied. The ratio of pentose to desoxypentose nucleic acid phosphorus also remained fairly constant over the interval at about 2.2. There was no significant difference in the amounts of P32 taken up over a period of 72 hr. by normal allantoic membranes and those infected with influenza virus, when inorganic radioactive phosphorus was placed in the allantoic sacs of 11-day embryonated eggs.

scholarly journals THE PROTEIN COATS OR "GHOSTS" OF COLIPHAGE T2

1958 ◽  
Vol 41 (5) ◽  
pp. 1067-1082 ◽  
Author(s):  
I. R. Lehman ◽  
Roger M. Herriott
Keyword(s):  
Nucleic Acid ◽  
Infected Cell ◽  
Inorganic Phosphate ◽  
Organic Phosphorus ◽  
Phosphorus Uptake ◽  
Inorganic Phosphorus ◽  
Ammonia Nitrogen ◽  
Normal Cells ◽  
E Coli ◽  
Infected Cells

Assimilation of oxygen, inorganic phosphate, and ammonia nitrogen by normal T2 phage and T2 ghost-infected E. coli B was studied. The rate of oxygen and phosphorus uptake by ghost-infected bacteria is similar to that of normal and phage-infected cells. The R.Q. in glucose-salts medium remains approximately 1. Assimilation of ammonia nitrogen by ghost-infected bacteria is maintained at a rate approximately 80 per cent of normal. The inorganic phosphate which is assimilated was found to be incorporated into TCA-soluble compounds which were rapidly released into the medium. Within 5 minutes after absorption of the ghosts there was a loss from the cell of TCA-soluble constituents including organic phosphorus and compounds which absorb at 260 mµ. No corresponding breakdown of nucleic acid present in the cell prior to infection could be detected. The incorporation of inorganic phosphate into organic linkages in the ghost-infected cell and its release into the medium were found to proceed at a rate approaching that of the incorporation of inorganic phosphorus into the nucleic acid of normal cells. The net increase in 260 mµ absorbing compounds appeared to be inhibited.

Phosphorylation of Histones in Normal Goose Erythrocytes

1971 ◽  
Vol 49 (9) ◽  
pp. 1062-1070 ◽  
Author(s):  
V. L. Seligy ◽  
J. M. Neelin
Keyword(s):  
Gene Expression ◽  
Nucleic Acid ◽  
Inorganic Phosphate ◽  
Specific Activity ◽  
Cell Maturation ◽  
Control Of Gene Expression ◽  
Nonhistone Protein ◽  
Free Nucleotide ◽  
And Control

Phosphate is taken up by erythrocytes in normal goose blood and incorporated into purified, characterized histones. The firmly bound radiophosphorus was not in the form of inorganic phosphate, free nucleotide, or phospholipid. That it was not due to contaminating nucleic acid or nonhistone protein was shown by chromatographic and electrophoretic localization.Most of the histone-bound radioactivity was recovered in slightly lysine-rich histone IIb1 (or f2a2) and in erythrocyte-specific histone V (or f2c), with the highest specific activity in the former. In these proteins the incorporated radiophosphorus was found only in phosphoserine.The differential incorporation did not correspond to alkali-labile phosphate contents, or to synthesis of the different proteins, and may reflect at least two different roles of phosphorylation in cell maturation and control of gene expression.

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