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At least 505 records · Page 28Linked to original sources
PubMed1969

Deoxyribonucleic acid-ribonucleic acid hybridization. Annealing and quantitative recovery of intact ribosomal ribonucleic acid molecules from hybrids.

A simple and efficient method for hybridization and subsequent recovery of non-fragmented ribosomal RNA from the hybrid is described. The procedure involves annealing of immobilized denatured DNA bound on cellulose nitrate membrane filters to complementary RNA in 50% (v/v) formamide-0.33m-potassium chloride-10mm-tris-hydrochloric acid buffer, pH7.4, at 33 degrees for 3hr. Under these conditions no detectable changes in the sedimentation coefficients of the input RNA were detected. The RNA can subsequently be recovered quantitatively from the hybrid in intact form by incubating the filters in formamide or in 85% (v/v) dimethyl sulphoxide. The applicability of the method for the evaluation of the absolute size of ribosomal RNA cistrons in Escherichia coli DNA and for the determination of the size of messenger RNA molecules is discussed.

Buffers↗
PubMed1969

Ribosomal RNA genes in bacteria: evidence for the nature of the physical linkage between 16S and 23S RNA genes in Bacillus subtilis.

The nature of the arrangement of 16S and 23S ribosomal RNA genes on the Bacillus subtilis chromosome was studied by means of a Cs(2)SO(4) density gradient centrifugation technique after complexing DNA-(ribosomal) RNA hybrids with mercuric ions. It was observed that the same fragments of single-stranded DNA with an average molecular weight of 1.9 x 10(6) which hybridize with 23S RNA also hybridize with 16S RNA. This indicates that genes for both these species of ribosomal RNA are physically linked on such DNA fragments. Some evidence suggests that the two types of genes are interspersed with each other, perhaps in sets, each of which contains a 16S and a 23S ribosomal RNA gene.

Bacillus subtilis↗
PubMed1970

Mechanism of viral carcinogenesis by DNA mammalian viruses. VII. Viral genes transcribed in adenovirus type 2 infected and transformed cells.

DNA-RNA hybridization-competition experiments were used to compare the virus-specific RNA sequences synthesized during productive infection with human adenovirus type 2 with those synthesized in virus-free adenovirus type 2 transformed cells. The "early" virus-specific RNA present at six hours after infection, prior to the onset of viral DNA synthesis, represents 8-20 percent (2 to 10 genes) of the viral genome. All viral RNA sequences synthesized early are also present "late," at 18 hours after infection. The base sequences transcribed in transformed cells are homologous to approximately 50 per cent of the sequences transcribed early after infection. Thus only 4 to 10 per cent of the viral genome, representing 1 to 5 viral genes, are transcribed in adenovirus type 2 transformed cells. The virus-specific RNA synthesized 18 hours after infection was not found in transformed cells, suggesting that either these late viral genes are not present or are not transcribed in adenovirus type 2 transformed cells.

Adenoviridae↗
PubMed1970

Genetic evolution of bacteriophage. II. Physical length of the homologous region in a hybrid between serologically and morphologically unrelated phages.

P22 and P221 DNA were labeled with (3)H-thymidine and hybridized with P22 DNA, P221 DNA, or Fels 1 DNA immobilized on Millipore membrane filters. With this technique, 31% of the P22 genome was computed to be homologous to the P221 genome. Since the molecular weight of P22 DNA is 2.65 x 10(7) daltons, the homologous region between P22 and P221 should be equivalent to about 8.2 x 10(6) daltons and contain 13,300 nucleotide pairs or about 4400 triplets. Moreover, 46% of the P221 genome was computed to be homologous to the P22 genome. Therefore, the molecular weight of P221 DNA was calculated to be about 1.80 x 10(7) daltons. Sucrose gradient centrifugation showed that the molecular weight of P221 DNA was 2.01 x 10(7) daltons. Thus, the P221 genome is about 25 to 30% smaller than the P22 genome. Further-more, 54% of the P221 genome was found to be homologous to the Fels 1 genome. However, the labeled P22 DNA hybridized with Fels 1 DNA was about 2% of the labeled P22 DNA hybridized with P22 DNA.

Carbon Isotopes↗
PubMed1975

Cloning, isolation, and characterization of replication regions of complex plasmid genomes.

EcoRI endonuclease-generated DNA fragments carrying replication regions of the F'lac and R6-5 plasmids have been cloned and isolated, using as a selection vehicle a nonreplicating ampicillin-resistance DNA fragment derived from a Staphylococcus aureus plasmid. Heteroduplex analysis of the constructed plasmid chimeras and the parent replicons has localized the cloned R6-5 replication region to a DNA segment between kilobase pair coordinates 1.0 and 88.0 on the R6-5 map. Physical proximity between the plasmid replication functions and the locus governing plasmid incompatibility has been shown for both parent replicons. The cloning method reported appears to be generally applicable for the identification and isolation of replication regions of a variety of complex genomes.

Centrifugation, Density Gradient↗