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J N Hansen

Publications and source records attributed to J N Hansen.

48 records · Page 3Linked to original sources

Hydroxyapatite chromatography of short single-stranded DNA.

Short single-stranded segments of calf thymus DNA were obtained by random cleavage with DNAase I. After treatment with various concentrations of DNAase I, fragment sizes were estimated using the ratio of total to terminal phosphorus. DNA populations ranging from 4--180 bases were obtained. Fragments with lengths up to 1140 were generated by shearing in a Virtis homogenizer. The hydroxyapatite elution profiles of sized populations were determined by elution with phosphate gradients. A curve relating elution molarities to single-strand chain length was 'biphasic', with the elution molarity being extremely sensitive to chain lengths below 50 nucleotides but much less sensitive to chain lengths above 100 nucleotides. These results show that single-stranded fragments below 50 nucleotides elute from hydroxyapatite appreciably before high molecular-weight denatured DNA using phosphate gradients. This is an important consideration when using hydroxyapatite to fractionate DNA populations which contain short single strands.

Animals↗

The specificity of S1 nuclease toward RNA-DNA hybrids as studied using isotopes of phosphorus-32 and phosphorus-33.

Hybrids were formed from Bacillus cereus DNA and ribosomal RNA. They were treated with various combination of S1 nuclease and ribonuclease, and the molar ratios of the RNA and DNA moieties remaining in the treated hybrids were determined using a 32P-33P dual-label technique. It was found that both S1 nuclease and ribonuclease are required to give hybrid with RNA and DNA in a perfect 1:1 molar ratio. It was noted that the dual-label technique which employs orthophosphate as the sole phosphorus source for both labels gives unambiguous molar ratios and obviates the need to calculate specific activities, make quench corrections, or correct for base content.

Bacillus cereus↗

Effect of some inhibitors derived from nitrite on marcomolecular synthesis in Bacillus cereus.

The effect of several inhibitors derived from nitrite on incorporation of [14C]uracil into ribonucleic acid of Bacillus cereus during outgrowth and vegetative growth has been determined. A heat-induced inhibitor presumably of the Perigo type was compared with the nitrosothiols of thioglycolate and beta-mercaptoethanol. All were found to effectively inhibit uracil incorporation at all stages tested. Phase-contrast microscopy revealed that inhibition of morphological events occurred either before germination or during early outgrowth, depending on inhibitor concentration. It was also found that a precursor to the heat-induced inhibitor could be isolated from dialyzable tryptone (a pancreatic digest of casein), which chromatographed as a single species on diethylaminoethyl-cellulose and Sephadex G-25, with an apparent molecular weight of about 2,000.

Anti-Bacterial Agents↗