Differential polyadenylation pattern of ovalbumin precursor RNAs during development.

The expression of the ovalbumin gene encoding for the major hen oviduct protein slows down with age. Analysis of Northern blots of electrophoretically separated total and poly(A) + RNA from oviducts of hens of different age with an ovalbumin-specific probe (nick-translated 9.5 kb ovalbumin gene DNA cloned into pBR322) revealed that the largest high molecular weight ovalbumin RNA precursor (7.9 kb band, representing the putative primary transcript of the ovalbumin gene) was most intense if total RNA from non-egg-laying old hen oviduct was checked as compared to that from egg-laying mature animals. On the other side, the 7.9 kb RNA precursor band was readily detected in the poly(A) + RNA from…

The determination of the DNA base composition in 19 species of adriatic sponges with high-pressure liquid cation-exchange chromatography.

Abstract The (adenine + thymine)/(guanine + cytosine) base ratios of 19 species of adriatic sponges have been determined by high-pressure liquid cation-exchange chromatography. The base ratios vary from 1.49 (Mycale massa) to 0.63 (Hippospongia communis) according to an (A+T) content of 59.7 and 38.6 mol%, respectively. The DNAs of sponges of the order Keratosa showed marked differences in their (A +T) contents (39.5 to 58.8 mol%) whereas those of Tetractinellida and Halichondrina were nearly identical (39.3 to 40.8 and 49.5 to 49.8 mol%, respectively). The 5-methylcytosine (5MC) content was determined in 8 sponge DNAs by a semiquantitative method. The values differed from 0.8 to 2.2 mol% o…

IV. An Improved Separation Method for Twenty two Compounds Related to Purine and 6-Thiopurine Metabolism Using High-Pressure Liquid Cation-Exchange Chromatography

Abstract An improved method is described for the separation of 22 compounds normally related to purine and 6-thiopurine metabolism in biological materials using high-pressure liquid cation-exchange chromatography on strongly acidic exchange resin. The column (0.18 × 100 cm) is eluted with 0.4 ᴍ ammonium formate, pH 4.6, at a linear flow velocity of 5.2 cm · min-1 at 50 °C. The elution volumes of sulphate anions, allopurinol, 6-thioxanthine, adenine, adenosine, and guanosine are demonstrated additionally to further 16 purine and 6-thiopurine compounds.

Thymine content of sea water as a measure of biosynthetic potential

A hydrolysis procedure along with a high-pressure liquid chromatographic procedure is given enabling simple and reliable thymine determinations in the nanogram range in different fractions of sea-water samples taken from three different locations in the Northern Adriatic Sea. The levels corresponded to 1–3 μg DNA per liter. From total polyanionic thymine, which had been precipitated as the cetyltrimethylammonium salt, the highest percentage was linked to the particulate fraction, with a definite subsurface minimum at 10 to 15 m. There was a corresponding maximum of a high molecular “non-particulate” thymine-containing fraction at the corresponding depth. From the bottom at 30 m upwards to a…

An isocratic high-pressure liquid chromatographic purification method for radioactively labeled deoxyribonucleoside triphosphates.

Abstract A method is described for the rapid purification of radioactively labeled deoxyribonucleoside tri­phosphates from their spontaneously emerging hydrolysis products deoxyribonucleoside diphosphate, deoxyribonucleoside monophosphate, and deoxyribonucleoside. The separations which are finished within 3 min or less are carried out on a 0.1X5 cm column filled with LiChrosorb-NH2 , using isocratic elution with 0.025 м potassium phosphate, pH 6 .8 , in a high-pressure liquid chromatograph at room temperature and a flow rate of 30 ml · h-1(flow velocity 63.7 cm·min-1).

A rapid separation of the four major deoxynucleosides and deoxyinosine by high-pressure liquid cation-exchange chromatography

The quantitative determination of metabolites of 6-mercaptopurine in biological materials. VII. Chemical synthesis by phosphorylation of 6-thioguanosine 5'-monophosphate, 5'-diphosphate and 5'-triphosphate, and their purification and identification by reversed-phase/ion-pair high-performance liquid chromatography and by various enzymatic assays.

Abstract A fast and reliable two-step method has been established for the chemical synthesis of 6-thioguanosine 5′-monophosphate, 6-thioguanosine 5′-diphosphate and 6-thioguanosine 5′-triphosphate starting from the ribonucleoside. In the first step, 6-thioguanosine dissolved in triethyl phosphate, at high yield reacts with phosphorus oxide trichloride to 6-thioguanosine 5′-monophosphate which is purified by anion-exchange chromatography on DEAE-Sephadex using a step gradient of hydrochloric acid. In the second step, 6-thioguanosine 5′-monophosphate dissolved in water, reacts with phosphoric acid in the presence of pyridine/dicyclohexyl carbodiimide and is converted to 6-thioguanosine 5′-dip…

Specificity of deoxyribonuclease hydrolysis determined by high-performance liquid anion-exchange chromatography

Characterization of Different Deoxyribonucleases in Human Lymphocytes

Abstract Deoxyribonucleases, Disc Electrophoresis, Lymphocytes Four groups of deoxyribonuclease activities from human lymphocytes have been characterized by deoxyribonuclease assay in DNA-containing polyacrylamide gels following their separation by disc-electrophoresis. All activities hydrolyse DNA endonucleolytically. One neutral deoxyribo­ nuclease found in the cytoplasmic fraction prefers native or UV-irradiated DNA over denatured DNA as substrate and is a 5′-monoester former. Two groups of acid deoxyribonuclease activities are detectable in the nuclear fraction. Both are 3′-monoester formers. One is as well active with denatured DNA as with native DNA, the other one shows the same activ…

Eine quantitative Bestimmungsmethode des Uracilgehaltes biologischen Materials im Nanogramm-Bereich mit Hilfe der Hochdruck-Flüssigchromatografie / A Method for the Quantitative Estimation of Uracil Content in Biological Materials in the Nanogramme-Range Using High-Pressure Liquid Chromatography

Abstract A method is described for the estimation of the uracil content in biological materials by means of high-pressure liquid chromatography. Hydrolysis of the tissues and total liberation of RNA bases are carried out in 70% perchloric acid. Less than 1 mg of the materials are needed for analysis. A pre-purification of the hydrolyzates is carried out by anion-exchange chromatography. Recoveries are estimated by isotope dilution analysis with [2-14C] labelled uracil. The method is highly sensitive - about 6000 pmol of uracil content can at least be estimated quantitatively - and analysis time is short. In routine analysis a single sample needs 4 hours to be completed. When preparing sever…

The use of high-pressure liquid cation-exchange chromatography for determination of the 5-methylcytosine content of DNA.