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J K Setlow

Publications and source records attributed to J K Setlow.

At least 55 records · Page 3Linked to original sources

Single-strand regions in the deoxyribonucleic acid of competent Haemophilus influenzae.

The deoxyribonucleic acid (DNA) of competent wild-type Haemophilus influenzae and rec1 mutant cells contains single-strand regions, as judged by alkaline sucrose sedimentation, benzoylated naphthoylated diethylaminoethyl-cellulose fractionation, and digestion with an enzyme specific for single-strand regions in DNA. In contrast, the DNA of competent rec2 cells does not contain single-strand regions. Since transforming DNA does not associate with recipient DNA in the rec2 mutant as it does in wild type and rec1, it is concluded that the single-strand regions in the DNA of the competent cells are important for an early step in recombination between cell DNA and transforming DNA.

Bromodeoxyuridine

Minicell production and bacteriophage superinducibility of thymidine-requiring strains of Haemophilus influenzae.

Aminopterin- or trimethoprin-resistant thymidine-requiring strains of Haemophilus influenzae produce minicells, and the ratio of minicells to cells increases during the stationary phase of growth. Strain LB11, isolated after mutagenesis of a thymidine-requiring strain (Rd thd), produces more minicells than the parent strain. The mutations involved in high frequency minicell production have been transferred into the wild type (strain Rd) by transformation. The thymidine requirement in the resulting strain, MCl, is essential for minicell production, since spontaneous revertants of MCl to prototrophy do not produce minicells. The ratio of minicells to cells was increased more than 10(3)-fold by differential centrifugation. The minicells contain little or no deoxyribonucleic acid (DNA). Phage HPlcl apparently cannot attach to minicells. Competent cells of LB11 and its thymidine-requiring parent strain produce defective phage as a result of exposure to transforming DNA, whereas only LB11 produces many defective phage in response to the competence regime alone. Competent HP1c1 and S2 lysogens of MC1 and Rd thd are also superinducible by transforming DNA, but competent LB11 lysogens produced about the same amount of HP1c1 or S2 phage with or without exposure to transforming DNA possibly because of competition between the induced defective phage and Hp1c1 or S2 phage.

Bacteriophages

Integration and repair of ultraviolet-irradiated transforming deoxyribonucleic acid in Haemophilus influenzae.

The extent of association between donor transforming deoxyribonucleic acid (DNA) and recipient DNA in Haemophilus influenzae as a function of ultraviolet (UV) dose to the transforming DNA has been measured by isopycnic analysis of lysates of (3)H-labeled recipient cells exposed to DNA labeled with (32)P and heavy isotopes. Except for doses above 15,000 ergs/mm(2), the results of these measurements are in good agreement with previous estimates made by another technique. Experiments with a mutant temperature sensitive for DNA synthesis and another mutant defective in excision of pyrimidine dimers suggest that the discrepancy between the methods of high doses results from DNA synthesis, in which portions of the associated donor DNA containing pyrimidine dimers are excised and broken down, and the components are reutilized for synthesis. Repair of UV-irradiated, transforming DNA during incubation of recipient cells is observed as an increase in transforming ability when fractions from CsCl gradients of cell lysates are assayed on excision-deficient cells. When transforming DNA containing markers of different UV sensitivities is used, repair of the UV-resistant nov marker by excision proficient cells takes place exclusively in the donor DNA that is associated with recipient DNA, and this repair is observed even in the absence of DNA synthesis. However, no repair is observed in the case of the more UV-sensitive str marker, possibly because excision events may remove a large fraction of the integrated str markers in addition to repairing a small fraction of the integrated DNA containing this marker.

Centrifugation, Density Gradient

Marker rescue in Haemophilus influenzae bacteriophage.

Rescue of wild-type markers from transfecting phage DNA in cómpetent Haemophilus influenzae cells by superinfection with temperature-sensitive phage (marker rescue) is approximately linearly dependent upon the concentration of transfecting DNA. The amount of marker rescue with a constant amount of transfecting DNA increases with increasing multiplicities of superinfecting phage up to about 4, and then decreases at higher multiplicities. Host restriction of transfecting DNA does not affect marker rescue. The frequency of wild-type recombinants from marker rescue is much greater than that from multiple infection with whole phages, and is comparable to that obtained with two mutant-transfecting DNAs. The amount of marker rescue decreases exponentially with time between entrance of the transfecting DNA and superinfection, and the rate of decrease is independent of map position of the rescued marker. Marker rescue is drastically reduced in the recombination-defective strains, rec1 and rec2.

Bacteriophages

Relationship between prophage induction and transformation in Haemophilus influenzae.

The interaction between transformation and prophages of HP1c1, S2, and a defective phage of Haemophilus influenzae has been investigated by measurement of (i) the effect of prophage on transformation frequency and (ii) the effect of transformation on phage induction. The presence of any of the prophages does not appreciably alter transformation frequencies in various Rec(+) and Rec(-) strains. However, exposure of competent lysogens to transforming deoxyribonucleic acid (DNA) may induce phage but only in Rec(+) strains, which are able to integrate transforming DNA into their genome. Transformation of Rec(+) lysogens with DNA irradiated with ultraviolet (UV) light causes the production of even more phage than results from unirradiated DNA, but this indirect UV induction is not as effective as direct induction by UV irradiation of lysogens. Both types of UV induction are influenced by the repair capacity of the host. Wild-type cells contain a prophage and can be induced by transformation to produce a defective phage, which kills a small fraction of the cells. Defective phage in wild-type cells are also induced by H. parainfluenzae DNA, and a much larger fraction of the cells is killed. Strain BC200, which is highly transformable but is not inducible for defective phage, is not killed by H. parainfluenzae DNA, suggesting that wild-type cells are killed by killed by this DNA because of phage induction. A minicell-producing mutant, LB11, has been isolated. Some phage induction occurs in this strain when the cells are made competent, unlike the wild type. A large majority of LB11 cells surviving the competence regime are killed by exposure to transforming DNA.

Bacteriophages

Bacteriophage of Haemophilus influenzae. 3. Morphology, DNA homology, and immunity properties of HPlcl, S2, and the defective bacteriophage from strain Rd.

The phages HP1c1 and S2 and a defective phage of Haemophilus influenzae have been compared. The morphology of the phages and the mol wt of their DNAs are similar, although the defective phage appears to have a different tail plate region. Electron microscope observation indicates that the defective phage does not attach to the cell surface, and its DNA appears to lack cohesive ends. The homology of the DNAs of the phages has been measured by hydridization. DNA from the defective phage shows little or no homology with the other phage DNAs. HP1c1 and S2 DNAs show a high level of homology. Each of these phages can form plaques on lawns of the lysogen of the other phage but at reduced plating efficiencies, suggesting that the two phages have related but not identical immunity systems.

Bacteriophages

Photoreactivation and gene dosage in yeast.

The amount of photoreactivating enzyme in tetraploid cells of Saccharomyces cerevisiae and the ability of the cells to be photoreactivated after ultraviolet irradiation are directly proportional to the number of genes per cell involved in the synthesis of photoreactivating enzyme.

Bacteriological Techniques