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Biomedical subjects

D Dubnau

Publications and source records attributed to D Dubnau.

108 records · Page 6Linked to original sources
PubMed1973

Fate of transforming deoxyribonucleic acid after uptake by competent Bacillus subtilis: phenotypic characterization of radiation-sensitive recombination-deficient mutants.

A collection of 16 isogenic recombination-deficient strains of Bacillus subtilis isolated on the basis of sensitivity to methyl methane sulfonate (MMS) or mitomycin C (MC) were characterized phenotypically. All were found to be somewhat sensitive to ultraviolet irradiation, MC, and MMS. The mutants were all blocked in "late" steps in the transformation process and were provisionally grouped into four categories on the basis of the various properties examined. Class I mutants were deficient in transformation and heterologous transduction with phage PBS1 but were transducible with homologous donors at nearly the wild-type frequency. They were blocked in donor-recipient complex (DRC) formation but formed essentially normal amounts of double-strand fragments (DSF) and single-strand fragments (SSF). The class IIa strain was deficient in transformation and PBS1 transduction, and formed DRC which was normal by all available physical and biological criteria. Class IIb mutants were deficient in transformation and PBS1 transduction, and failed to form DRC. They did produce DSF and SSF. Class III mutants were deficient in transformation, were normal in PBS1 transduction, and formed DRC which was physically indistinguishable from that of the Rec(+) parent although with slightly lowered donor-type transforming activity. Class IV strains were deficient in PBS1 transduction but were transformed at nearly the wild-type efficiency. None of the mutant strains was deficient in the adenosine triphosphate-dependent deoxyribonuclease.

Alkanesulfonates↗
PubMed1973

Conditions affecting the isolation from transformed cells of Bacillus subtilis of high-molecular-weight single-stranded deoxyribonucleic acid of donor origin.

The deoxyribonucleic acid (DNA) extraction procedure of Piechowska and Fox was evaluated to determine which steps are required for the isolation of high-molecular-weight single-stranded material from transformed cultures of Bacillus subtilis. The results indicate that high-molecular-weight single-stranded DNA can be isolated when certain basic proteins are present at the time of lysis. In the absence of such protective agents as lysozyme or cytC the single-stranded DNA is degraded. The single-stranded DNA can also be protected by being treated with lysozyme at low temperature. The high molecular weight of this single-stranded material and its kinetics of appearance are consistent with its being an intermediate in the transformation process.

Bacillus subtilis↗
PubMed1973

Kinetic analysis of the products of donor deoxyribonucleate in transformed cells of Bacillus subtilis.

This paper describes the major transmutations of donor deoxyribonucleic acid (DNA) after uptake by competent Bacillus subtilis cells. Kinetic experiments confirm that after exposure to competent cells, donor DNA is converted to double-stranded fragments (DSF) which can be isolated as early as 30 s from the beginning of the reaction. At this time, DSF represent the only identifiable product of donor origin. After 1 to 2 min, DSF are converted to deoxyribonuclease-resistant forms, identified as single-stranded DNA fragments (SSF). SSF are intermediates in the transformation process leading to the formation of donor-recipient complex. This component makes its appearance between 2 to 4 min from the beginning of the transformation process. All the donor-recipient complexes found at the end of the reaction can be accounted for quantitatively by the DSF and the SSF found in the initial stages of transformation. A quantitative discussion of the transformation process is included.

Bacillus subtilis↗
PubMed1972

Fate of transforming deoxyribonucleic acid after uptake by competent Bacillus subtilis: size and distribution of the integrated donor segments.

A series of sheared donor-recipient complex samples were prepared from a competent Bacillus subtilis culture which had been transformed by using (3)H, (2)H-labeled deoxyribonucleic acid. Measurement of the molecular weights and buoyant densities of these samples led to the following conclusions. (i) The average size of the single-stranded donor segment which is integrated is 2.8 x 10(6). (ii) These donor segments are not randomly distributed in the recipient genome, but are clustered in groups, each group containing several discrete donor segments.

Bacillus subtilis↗
PubMed1971

Fate of transforming DNA after uptake by competent Bacillus subtilis: failure of donor DNA to replicate in a recombination-deficient recipient.

The fate of radioactively-labeled transforming DNA was studied in a recombination-deficient strain of Bacillus subtilis that carried the recB2 mutation (Rec(-)) and was sensitive to radiation. Experiments performed with extracts of this strain after transformation showed that the recovery of donor transforming activity and the appearance of recombinant transforming activity occurred to the same extent as in the Rec(+) strain. Sucrose gradient analyses revealed that donor-recipient complex is also formed to the same extent in the Rec(-) and Rec(+) strains, but that 80-90% of the donor genetic material in the complex failed to replicate in the Rec(-) mutant.

Bacillus subtilis↗
PubMed1970

Genetic mapping of antibiotic resistance in markers Bacillus subtilis.

Several antibiotic resistance markers in Bacillus subtilis have been mapped by three-factor transduction crosses using bacteriophage PBS1. These markers, which are thought to be located in genes coding for ribosomal proteins, are within a segment comprising about 5 per cent of the B. subtilis genetic map, close to the major group of rRNA cistrons. The order of genetic makers in this region was found to be cysA14 mic-1 dagger str-1 (ery-1, ole-2) spc-2 bry-2 lin-2. The positions of neo-2, nea-1, and kan-2 are uncertain, although they are located in this region to the right of cysA14.

Anti-Bacterial Agents↗
PubMedDate not supplied

Internalizing DNA.

The steps involved in the transformation of Bacillus subtilis are reviewed. These include the initial binding, processing and passage of DNA across the cell wall and transport across the plasma membrane. Our understanding of the roles of the proteins known to be required for these steps is reviewed.

Bacillus subtilis↗