Analysis of mutations at the adenine phosphoribosyltransferase gene locus in wild-type and thymidine kinase-deficient Friend cells

1996 ◽  
Vol 24 (1) ◽  
pp. 105S-105S ◽  
Author(s):  
Paula L. Hyland ◽  
Mark W. McKinney ◽  
Anne. L. Keegan ◽  
Patrick G. McKenna ◽  
Yvonne A. Barnett
Keyword(s):  
Thymidine Kinase ◽  
Gene Locus ◽  
Wild Type ◽  
Adenine Phosphoribosyltransferase ◽  
Friend Cells

Sequence analysis of spontaneously-arising mutations at the aprt locus in wild-type and thymidine kinase-deficient Friend cells: Evidence for strand slippage-misalignment mechanism in formation of deletions

1997 ◽  
Vol 25 (1) ◽  
pp. 127S-127S ◽  
Author(s):  
PAULA L. HYLAND ◽  
MARK W. MCKINNEY ◽  
ANNE L. KEEGAN ◽  
PATRICK. G. MCKENNA ◽  
MARTIN. D. CURRAN ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Thymidine Kinase ◽  
Wild Type ◽  
Friend Cells

Developmentally controlled telomere addition in wild-type and mutant paramecia

1988 ◽  
Vol 8 (1) ◽  
pp. 251-258
Author(s):  
J D Forney ◽  
E H Blackburn
Keyword(s):  
Surface Antigen ◽  
Base Pair ◽  
Gene Locus ◽  
Gene Sequence ◽  
Genetic Locus ◽  
Single Site ◽  
Paramecium Tetraurelia ◽  
Wild Type ◽  
Telomeric Sequences ◽  
Antigen Gene

We analyzed sites of macronuclear telomere addition at a single genetic locus in Paramecium tetraurelia. We showed that in homozygous wild-type cells, differential genomic processing during macronuclear development resulted in the A surface antigen gene being located 8, 13, or 26 kilobases upstream from a macronuclear telomere. We describe variable rearrangements that occurred at the telomere 8 kilobases from the A gene. A mutant (d48) that forms a telomere near the 5' end of the A gene was also analyzed. This mutant was shown to create simple terminal deletions; telomeric repeats were added directly to the truncated wild-type A gene sequence. In both the mutant and wild-type cells, the telomeric sequences (a mixture of C4A2 and C3A3 repeats) were added to various sequences within a specific 200- to 500-base-pair region rather than to a single site. No similarities were found in the primary sequences surrounding the telomere addition sites. The mutation in d48 changed the region of telomere addition at the A gene locus; this is the first example in ciliates of a mutation that affects the site of telomere addition.

Human Thymidine Kinase Gene Locus: Assignment to Chromosome 17 in a Hybrid of Man and Mouse Cells

Science ◽  
1971 ◽  
Vol 173 (3993) ◽  
pp. 244-245 ◽  
Author(s):  
O. J. Miller ◽  
P. W. Allderdice ◽  
D. A. Miller ◽  
W. R. Breg ◽  
B. R. Migeon
Keyword(s):  
Thymidine Kinase ◽  
Gene Locus ◽  
Chromosome 17 ◽  
Thymidine Kinase Gene ◽  
Kinase Gene ◽  
Mouse Cells

scholarly journals Location of sequences in polyomavirus DNA that are required for early gene expression in vivo and in vitro.

1984 ◽  
Vol 4 (12) ◽  
pp. 2594-2609 ◽  
Author(s):  
C R Mueller ◽  
A M Mes-Masson ◽  
M Bouvier ◽  
J A Hassell
Keyword(s):  
Gene Expression ◽  
Thymidine Kinase ◽  
Dna Sequences ◽  
Viral Genome ◽  
Simian Virus 40 ◽  
Early Gene ◽  
Wild Type ◽  
Transcription In Vitro

To define the DNA sequences required for the expression of the polyomavirus early transcription unit, we cloned part of the viral genome in a plasmid vector, isolated mutants bearing lesions introduced in vitro within DNA sequences upstream of the transcriptional start site, and measured the capacity of these various mutant genomes to transform cells and to function as templates for transcription in vitro by comparison with wild-type DNA. One set of mutants bore 5' unidirectional deletions beginning at position -810 and extending downstream to position +4. Another set of mutants bore 3' undirectional deletions starting at position +4 and progressing upstream to position -311. The last set of mutants bore internal deletions between positions -810 and +4. Analyses of the properties of these mutant DNAs led us to conclude that the region between positions -403 and -311 includes an enhancer of gene expression. Deletion of this area from the viral genome reduced gene expression in vivo to 1 to 2% of wild-type levels, as measured by transformation assays. Moreover, this region increased the frequency of transformation of thymidine kinase-negative Rat-2 cells by the herpes simplex virus thymidine kinase (tk) gene from 5- to 20-fold. This occurred only if the polyomavirus sequences were covalently linked to the tk gene and then occurred independently of their orientation or position relative to the tk gene. A second transcriptional element is located downstream of the enhancer between positions -311 and -213. This element together with the enhancer was sufficient to bring about transformation of Rat-1 cells at nearly wild-type frequencies, and together these elements constitute the minimal sequences required for gene expression in vivo. The sequences making up the second element may be functionally duplicated downstream of position -165 (between positions -165 and -60). This was revealed by the characterization of mutant genomes with deletions between positions -349 and -60. The role of these redundant elements is not known; however, they may be analogous to the 21-base-pair repeats of simian virus 40. Finally, sequences between positions -57 and -1 were required for accurate and efficient transcription in vitro. However, this DNA stretch, which includes the TATA box and major transcriptional start sites, was not absolutely required for gene expression in vivo. We conclude that the polyomavirus promoter comprises multiple functional elements which are distributed across a DNA stretch of about 400 base pairs.

scholarly journals Cloning of the Koi Herpesvirus Genome as an Infectious Bacterial Artificial Chromosome Demonstrates That Disruption of the Thymidine Kinase Locus Induces Partial Attenuation in Cyprinus carpio koi

2008 ◽  
Vol 82 (10) ◽  
pp. 4955-4964 ◽  
Author(s):  
B. Costes ◽  
G. Fournier ◽  
B. Michel ◽  
C. Delforge ◽  
V. Stalin Raj ◽  
...  
Keyword(s):  
Bacterial Artificial Chromosome ◽  
Thymidine Kinase ◽  
Artificial Chromosome ◽  
Wild Type ◽  
Bac Clone ◽  
Recombinant Viruses ◽  
Koi Herpesvirus ◽  
Cyprinus Carpio Koi

ABSTRACT Koi herpesvirus (KHV) is the causative agent of a lethal disease in koi and common carp. In the present study, we describe the cloning of the KHV genome as a stable and infectious bacterial artificial chromosome (BAC) clone that can be used to produce KHV recombinant strains. This goal was achieved by the insertion of a loxP-flanked BAC cassette into the thymidine kinase (TK) locus. This insertion led to a BAC plasmid that was stably maintained in bacteria and was able to regenerate virions when permissive cells were transfected with the plasmid. Reconstituted virions free of the BAC cassette but carrying a disrupted TK locus (the FL BAC-excised strain) were produced by the transfection of Cre recombinase-expressing cells with the BAC. Similarly, virions with a wild-type revertant TK sequence (the FL BAC revertant strain) were produced by the cotransfection of cells with the BAC and a DNA fragment encoding the wild-type TK sequence. Reconstituted recombinant viruses were compared to the wild-type parental virus in vitro and in vivo. The FL BAC revertant strain and the FL BAC-excised strain replicated comparably to the parental FL strain. The FL BAC revertant strain induced KHV infection in koi carp that was indistinguishable from that induced by the parental strain, while the FL BAC-excised strain exhibited a partially attenuated phenotype. Finally, the usefulness of the KHV BAC for recombination studies was demonstrated by the production of an ORF16-deleted strain by using prokaryotic recombination technology. The availability of the KHV BAC is an important advance that will allow the study of viral genes involved in KHV pathogenesis, as well as the production of attenuated recombinant candidate vaccines.

scholarly journals Competition and complementation between thymidine kinase-negative and wild-type herpes simplex virus during co-infection of mouse trigeminal ganglia

2006 ◽  
Vol 87 (12) ◽  
pp. 3495-3502 ◽  
Author(s):  
Shih-Heng Chen ◽  
Yu-Wen Lin ◽  
Anthony Griffiths ◽  
Wen-Yen Huang ◽  
Shun-Hua Chen
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Thymidine Kinase ◽  
Antiviral Drug ◽  
Wild Type Virus ◽  
Trigeminal Ganglia ◽  
Wild Type ◽  
Viral Genomes ◽  
Simplex Virus

Laboratory strains of herpes simplex virus lacking thymidine kinase (TK) cannot replicate acutely to detectable levels in mouse trigeminal ganglia and do not reactivate from latency. However, many pathogenic clinical isolates that are resistant to the antiviral drug acyclovir are heterogeneous populations of TK-negative (TK−) and TK-positive (TK+) viruses. To recapitulate this in vivo, mice were infected with mixtures of wild-type virus and a recombinant TK− mutant in various ratios. Following co-infection, the replication, number of latent viral genomes and reactivation efficiency of TK+ virus in trigeminal ganglia were reduced in a manner related to the amount of TK− virus in the inoculum. TK+ virus did not always complement the acute replication or increase the number of latent viral genomes of TK− mutant in mouse ganglia. Even so, TK+ virus could still confer the pathogenic phenotype to a TK− mutant, somehow providing sufficient TK activity in trans to permit a TK− mutant to reactivate from latently infected ganglia.

scholarly journals Increased sensitivity to cell killing and mutagenesis by chemical mutagens in thymidine-kinase-deficient subclones of a Friend murine leukaemia cell line

1982 ◽  
Vol 40 (2) ◽  
pp. 207-212 ◽  
Author(s):  
P. G. McKenna ◽  
A. A. Yasseen
Keyword(s):  
Thymidine Kinase ◽  
Wild Type ◽  
Ethyl Methane Sulphonate ◽  
Leukaemia Cell ◽  
Leukaemia Cell Line ◽  
Ethyl Methane ◽  
Chemical Mutagens ◽  
Methane Sulphonate ◽  
Increased Sensitivity ◽  
Unit Dose

SUMMARYWild-type Friend murine leukaemia (clone 707) cells and two thymidinekinase-deficient subclones, 707BUE and 707BUF, were compared for sensitivity to killing and mutagenesis by the chemical mutagens, ethyl methane sulphonate (EMS),N-methyl-N′-nitro-N-nitrosoguanidine (MNNG), mitomycin C (MMC), and methyl methane sulphonate (MMS). The two thymidine-kinase-deficient subclones were significantly more sensitive to killing by each of the four chemical mutagens than were wild-type cells. The increased sensitivity to killing by the four mutagens was also reflected in increased mutagenesis (per unit dose of mutagen) to 6-thioguanine resistance. In the light of these results, the significance of thymidine kinase in DNA repair and mutagenesis is discussed.

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