Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.1.21 (thymidine kinase)
 7,561 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have analysed Epstein-Barr virus (EBV)- and herpes simplex virus (HSV)-infected cells for evidence of antigenic conservation of virus-coded proteins. Immunofluorescence and Western blot analyses of EBV-transformed cell lines demonstrated the presence of proteins that are antigenically related to the HSV alkaline DNase, infected cell-specific protein 34/35, glycoprotein B, thymidine kinase and the major DNA-binding protein. These proteins were characterized on the basis of Mr and possible kinetic class.
J Gen Virol 1988 Aug
PMID:Immunological conservation between Epstein-Barr virus and herpes simplex virus. 284 14

The morphologically normal rat fibroblast cell line Rat-2 was used as a target cell type to test the transforming ability of a human papillomavirus (HPV-1a). To this end, molecularly cloned HPV-1a genomes were introduced into cultured Rat-2 cells in cotransfection experiments using a cloned herpes simplex virus thymidine kinase gene as a selectable phenotypic maker. In each of 13 HPV-1a-positive cell clones examined the papillomavirus DNA sequences were associated with the high molecular weight fraction of the cellular DNA, and restriction endonuclease plus Southern blotting analyses revealed patterns of hybridization which were consistent with integration of the viral genomes. Even Rat-2 clones containing multiple copies of the entire HPV-1a genome retained the normal, i.e. flat, cell morphology and were unable to grow in soft agar. De novo methylation of the HPV-1a sequences in many Rat-2 cell clones was evidenced by resistance of the viral DNA to complete cleavage with the HpaII endonuclease. Two out of three cell lines harbouring multiple copies of the HPV-1a genome contained detectable levels of HPV-1a transcripts, whereas no transcripts were detected in the third such cell line in which the viral HpaII sites were methylated virtually to completion. These results are consistent with the notion that HPV-1a genes are expressed inefficiently in Rat-2 cells; consequently integration of the viral DNA occurs, and there is no effect of the virus on the growth properties of this cell type. It is possible that methylation of the HPV-1a sequences is responsible for the low levels of expression of the viral genome.
J Gen Virol 1985 May
PMID:Introduction of cloned human papillomavirus 1a DNA into rat fibroblasts: integration, de novo methylation and absence of cellular morphological transformation. 298 96

Human cell lines that contain and express the gene encoding the adenovirus type 5 DNA-binding protein (Ad5 DBP) are very useful for the isolation of adenovirus mutants with an altered DBP. In order to obtain these cells, human 143 tk- cells were transfected, using the calcium phosphate technique, with plasmids containing the Ad5 DBP gene and the herpes simplex virus thymidine kinase (HSV tk) gene as a selectable marker. Characterization of several tk+ transformants revealed that these cells did contain the HSV tk gene, but in none of these cells could Ad5 DBP DNA sequences be detected. However, when 143 tk- cells were co-transfected with a plasmid containing the Ad5 DBP gene and another plasmid carrying early region E1, integration of the Ad5 DBP gene in chromosomal DNA could be detected. Integration of Ad5 DNA sequences was also observed when transfection was performed with plasmids containing the Ad5 DBP gene and the long terminal repeat of Moloney murine leukaemia virus. By employing a radioimmunoassay it could be shown that DBP-related proteins were synthesized in two of the cell lines containing the Ad5 DBP gene. Since both cell lines support the growth of the temperature-sensitive viral DBP mutant, H5ts125, at the non-permissive temperature, the DBP-related proteins expressed in these cells must be functional.
J Gen Virol 1985 Nov
PMID:Transformation of human 143 tk- cells with plasmids containing the gene encoding the adenovirus DNA-binding protein. 299 73

Intravenous inoculation of 4-week-old female NIH (inbred) mice with herpes simplex virus type 1 (HSV-1) strain P2C6 (defective in thymidine kinase) produced bilateral hind limb paralysis in nearly all animals by the 5th day after inoculation; very few mice died. In male mice the incidence and severity of paralysis was considerably lower than in females. The parental strain, CL(101), produced similar paralysis but all mice died by day 7. Observations on paralysis and death after intravenous inoculation are given for other strains of HSV-1 and HSV-2. By day 1 after inoculation of P2C6 significant virus replication had occurred in the adrenal glands but in none of the other organs tested. Titres of virus were similar in the adrenal glands of male and female mice. Histology of the adrenals showed most extensive replication in the cortex with some involvement of the medulla, particularly at the corticomedullary junction. By the 2nd and 3rd days, virus was detected in the lower thoracic spinal cord of both male and female animals but clearance was possibly quicker from males. Adrenalectomy proved that virus reached the cord via the adrenals. In the cord the infection was associated with bilateral demyelination in the ventral white matter as early as day 3.
J Gen Virol 1986 Feb
PMID:Infection of the adrenal gland as a route to the central nervous system after viraemia with herpes simplex virus in the mouse. 300 39

We have established procedures for reisolating a transfected gene from mammalian cells by selection in Escherichia coli for the function of the gene product using the Herpes simplex virus thymidine kinase gene as a model. Rescue of the gene is accomplished by three different methods. The tk gene is recloned into a plasmid in which it is hooked up by either the lac promoter or a lac/tk hybrid promoter, or the original plasmid is cut out of the host cell DNA. As the lac/tk hybrid gene can be expressed and selected both in the mammalian and E. coli cells, this type of gene rescue allows investigations on mutagenesis and methylation processes. Additionally, it offers a simple way of studying the integration of the transfected gene into the mammalian genome.
Mol Gen Genet 1985
PMID:Rescue of transfected genes from mammalian cells by functional selection in Escherichia coli. 300 30

The nucleotide sequence of the coding region of the thymidine kinase gene from each of three mutant strains of herpes simplex virus type 1 and from the parental strain, SC16, has been determined. The mutants were known to express thymidine kinase enzymes with distinct substrate binding properties. Consideration of the lesions in the genes responsible for these altered biochemical properties. Consideration of the lesions in the genes responsible for these altered biochemical properties has led us to postulate a preliminary model for the active centre of the enzyme, involving the cooperation of three distinct regions of the polypeptide.
J Gen Virol 1986 Apr
PMID:Evidence that the 'active centre' of the herpes simplex virus thymidine kinase involves an interaction between three distinct regions of the polypeptide. 300 62

Using vaccinia virus as a selection and cloning vehicle, a thymidine kinase (TK) gene of fowlpox virus (FPV) has been identified. A plasmid, pF130, containing part of the HindIII-F region of vaccinia virus was used to shotgun clone EcoRI fragments of FPV DNA into TK- vaccinia virus and select for TK+ recombinants. The TK+ recombinant vaccinia virus contained a 5.5 kb EcoRI fragment of FPV. This FPV fragment was cloned into pUC9 and the presence of the TK gene in this fragment was confirmed by its ability to rescue TK+ vaccinia virus from TK- virus, when inserted into pF130. A recombinant vaccinia virus containing this FPV fragment induced TK enzyme activity in the cytoplasm of infected cells. The vaccinia virus RNA polymerase appeared able to recognize the FPV promoter sequences of the FPV TK gene since the fragment operated in the marker rescue, irrespective of its orientation to the vaccinia virus promoter in pF130. Using restriction enzyme analysis, insertion of subfragments of the 5.5 kb FPV fragment into pF130 and marker rescue, we were able to map the position of the TK gene in the 5.5 kb EcoRI fragment. This approach may facilitate identification and cloning of TK genes from other poxviruses.
J Gen Virol 1986 Aug
PMID:Identification and cloning of the fowlpox virus thymidine kinase gene using vaccinia virus. 301 54

Previously, mouse zygotes were microinjected with the recombinant plasmid pMA3, which contains the Herpes simplex virus thymidine kinase gene attached to the promoter region of the Rous sarcoma virus (Gazaryan et al. 1984a). In the present work the pMA3 fragment with the flanking genomic sequences was isolated from the DNA of one transgenic Fo mouse by the "plasmid rescue" technique. The rescued plasmid (pMAR1) lacked all virus-specific sequences and retained only some pBR322 sequences. The flanking region at one end of the integrated pBR322-specific fragment contained a highly conserved mouse repetitive sequence. The possible mechanisms of rearrangement of foreign DNA in germ line cells are discussed.
Mol Gen Genet 1986 May
PMID:Rearrangements of microinjected recombinant DNA in the genome of transgenic mice. 301 83

This communication demonstrates the usefulness of the plasmid rescue procedure for recovery of plasmids from transgenic mice. We have microinjected the plasmid pSK1 harbouring the Herpes simplex virus thymidine kinase gene into fertilized mouse oocytes and succeeded in recovering plasmids from newborns by transformation of E. coli either with HindIII cut cellular DNA or with uncut DNA. The majority of the rescued plasmids were indistinguishable from pSK1 by restriction analysis. The rescued plasmids proved to be functionally active in a transient expression assay in mouse Ltk- cells. The pSK1 DNA sequences were inherited by up to 90% of the second generation progeny mice, which is not in agreement with a Mendelian transmission of heterozygous markers integrated into a single site of the chromosome. These data support the assumption that germ line transmission of non-integrated episomal plasmids can occur.
Mol Gen Genet 1986 Aug
PMID:Rescue of a tk-plasmid from transgenic mice reveals its episomal transmission. 302 Mar 70

HeLa cells generally do not respond well to interferon (IFN). We have used is-1, an IFN-sensitive mutant of mengovirus, to select a clone of IFN-responsive HeLa cells (F-H12). At moderate levels of human alpha/beta IFN, is-1 yields were fivefold lower in these cells than in similarly protected control cells. In contrast, wild-type mengovirus, vesicular stomatitis virus and a wild-type and thymidine kinase-negative strains of herpes simplex virus type 1 grew equally well in both cell lines. By a cell survival assay, the F-H12 line was up to 100 times more responsive to IFN than the parental line when challenged by is-1. 2'-5'-Oligo(A)-dependent endonuclease activity was the same in both lines. These observations cannot be accounted for by enhanced induction of IFN following infection.
J Gen Virol 1986 Nov
PMID:Selection and characterization of an interferon-responsive clonal cell line of HeLa cells. 302 28


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