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Open AccessJournal ArticleDOI

Integration of the Deoxyribonucleic Acid of Adenovirus Type 12 into the Deoxyribonucleic Acid of Baby Hamster Kidney Cells

Walter Doerfler
- 01 Nov 1970 - 
- Vol. 6, Iss: 5, pp 652-666
TLDR
Evidence was presented that the deoxyribonucleic acid (DNA) of adenovirus type 12 (Ad12) is integrated by covalent linkage into the DNA of baby hamster kidney cells (BHK-21 cells).
Abstract
In a previous report, evidence was presented that the deoxyribonucleic acid (DNA) of adenovirus type 12 (Ad12) is integrated by covalent linkage into the DNA of baby hamster kidney cells (BHK-21 cells). These studies have been extended. The DNA of Ad12 and that of BHK-21 cells grown in medium containing 5-bromodeoxyuridine could be separated by equilibrium centrifugation in alkaline CsCl density gradients. BHK-21 cells were infected with 3H-labeled Ad12, and the total intracellular DNA was analyzed at various times after infection in alkaline CsCl density gradients. The 3H label in the position of cellular DNA hybridized predominantly with viral DNA and to a lesser extent also with cellular DNA. Replication of viral DNA could not be detected in BHK-21 cells. The appearance of viral 3H label in the density stratum of cellular DNA was not significantly affected when DNA synthesis in Ad12-infected BHK-21 cells was inhibited >96% by cytosine arabinoside. These findings provided additional evidence for integration of Ad12 DNA into the DNA of BHK-21 cells. It could be calculated that 5 to 55 Ad12 DNA equivalents per cell are integrated. Replication of viral or cellular DNA was not required for integration. Inhibition of protein or ribonucleic acid synthesis interfered with integration only slightly.

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Citations
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Detection of Epstein-Barr Viral Genome in Nonproductive Cells

TL;DR: Hybridization experiments have shown that EB viral DNA comprises between 0.06 and 1.6% of the total DNA of Burkitt lymphoma cells and established leucocyte cell lines.
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DNA methylation--a regulatory signal in eukaryotic gene expression

TL;DR: The DNA of prokaryotes contains the modified bases N 6-methyl-adenine (6-mA) and 5- methyl-cytosine (5-mC); other modified bases have apparently not been found.
Journal ArticleDOI

Patterns of integration of viral dna sequences in the genomes of adenovirus type 12-transformed hamster cells

TL;DR: Analysis of the integration patterns of Ad12 DNA in the four hamster lines investigated reveals that some of the viral DNA molecules are fragmented prior to or during integration, which is compatible with, but does not prove, the notion that at least a portion of theiral DNA sequences, is integrated into repetitive sequences, or else that the integrated viral sequences have been amplified after integration.
Journal ArticleDOI

Transcription of the genome of adenovirus type 12. III. Maps of stable RNA from productively infected human cells and abortively infected and transformed hamster cells.

TL;DR: The results of the mapping experiments demonstrate that the mRNA's isolated early from productively and abortively infected and from two lines of transformed cells are derived from the same or similar regions of the adenovirus type 12 genome.
Journal ArticleDOI

The response of BHK21 cells to infection with type 12 adenovirus: II. Relationship of virus-stimulated DNA synthesis to other viral functions☆

TL;DR: The results are taken to indicate that the virus initiates chromosomal DNA synthesis but that infection eventually results in extensive destruction of the cell genome, and thus the death of the cells.
References
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Journal ArticleDOI

A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid

TL;DR: The present study arose from the observation that a more intense colour was sometimes produced if, instead of being heated at 1000 for 10 min., the reaction mixture was allowed to stand overnight at room temperature.
Journal ArticleDOI

Plaque formation and isolation of pure lines with poliomyelitis viruses.

TL;DR: Pure virus lines were established by isolating the virus population produced in single plaques, which had the same morphological, serological, and pathogenic properties as the parent strain.
Journal ArticleDOI

A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane

TL;DR: An improved method for the formation of DNA—RNA hybrids is described, quantitatively more certain than annealing in liquid, since the competing DNA renaturation reaction is suppressed.
Journal ArticleDOI

A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells.

TL;DR: The covalently closed circular duplex DNA structure, originally identified in polyoma viral DNA,1’2 has been assigned to the mitochondrial DNA’s in ox and sheep heart, in mouse and chicken liver, and in unfertilized sea urchin egg.
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