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Thymidine Kinase

Thymidine kinase is an essential enzyme involved in DNA synthesis and repair.
It catalyzes the phosphorylation of thymidine to thymidine monophosphate, a key step in the salvage pathway of pyrimidine nucleotide biosynthesis.
This enzyme plays a crucial role in cell proliferation and is a marker for cellular growth and division.
Thymidine kinase has been extensively studied in the context of cancer, viral infections, and gene therapy, making it an important target for research and therapeutic development.
PubCompare.ai's AI-powered platform can help optimize your Thymidine kinase research by providing access to the best protocols, pre-prints, and patents, enhacing reproducibility and accuracy through intelligent analysis to identify the most effective methods and products.
Leverage PubComapre.ai to take your Thymidine kinase research to the next level.

Most cited protocols related to «Thymidine Kinase»

1
Genetic Manipulation of Bacteroides thetaiotaomicron
Cited 80 times

Protocol full text hidden due to copyright restrictions

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Publication 2008
Agar Alleles Bacteria Bacteroides Blood Cloning Vectors Deletion Mutation Deoxyuridine Erythromycin Gene Deletion Genes Genetic Vectors Genome Intergenic Region Nucleotides Oligonucleotide Primers Parent Plasmids Serine-Specific tRNA Strains Thymidine Kinase
2
Automated Docking of Diverse Protein Structures
Cited 43 times
For each target, we assembled all
UniProt accession codes (species) with any raw ChEMBL compounds (ligands,
decoys, marginal ligands, or marginal decoys). For only those accession
codes, structures were extracted using the ChEMBL to PDB mapping,
except P07700 was manually added to ADRB1 to include six more rare
structures for that GPCR. This procedure neglects those PDB structures
that belong to an accession code having no ChEMBL compounds. For example, 1KIM is the PDB structure
of thymidine kinase (KITH) in the original DUD. This KITH structure
is from herpes virus (UniProt P03176), an accession code with no raw
compounds extracted from ChEMBL, and is thus not included in the ChEMBL/PDB
intersection used to construct the new DUD. Still, 5025 PDB codes
were sent to an updated DOCK Blaster pipeline for automated docking
preparation (Supporting Information Figure S1D). In some cases, an unambiguous ligand could not be found to indicate
the binding site, but we were able to assign 565 additional ligands
by manually inspecting over 1300 structures. Ultimately, 3692 structures
completed input grid preparation, and all but two finished docking
and enrichment analysis. Clustered ligands sets were docked to property-matched
decoys (both described below) using ECFP4 fingerprints and removing
the most similar 75% of queried decoys. DOCK 3.6 was run using SEV
ligand desolvation (as below). For each target, enrichment, resolution,
and organism were collected and sorted by enrichment in pdb_analyze.txt,
available online at http://dude.docking.org. Crude notes
on the selection process are recorded in pdb_selection.txt, and the
picked structure is listed in pdb_blessed.txt. AA2AR and DRD3 docking
preparations were provided by Jens Carlson,44 (link),45 CXCR4 partially by Dahlia Weiss,3 (link) ADRB1
by Peter Kolb (personal communication), and AMPC by Sarah Barelier,
Oliv Eidam, and Inbar Fish (unpublished results).
Mysinger M.M., Carchia M., Irwin J.J, & Shoichet B.K. (2012). Directory of Useful Decoys, Enhanced (DUD-E): Better Ligands and Decoys for Better Benchmarking. Journal of Medicinal Chemistry, 55(14), 6582-6594.
Publication 2012
ADRB1 protein, human Binding Sites CXCR4 protein, human Dahlia Fishes Ligands Rumex Simplexvirus Thymidine Kinase
3
Generation and Characterization of IL-15 Deficient Mice
Cited 30 times
C57BL/6 mice genetically deficient in IL-15 were generated by homologous recombination in embryonic stem (ES) cells. The structure of the murine IL-15 genomic locus has been described 21. A gene targeting vector was constructed in which a 7.5-kb SpeI-EcoRV fragment containing IL-15 exons 3–5, encoding amino acids 1–65 of the primary translation product, was replaced with a PGK-neo cassette. A thymidine kinase cassette (MC-TK) was inserted into the 5′ end of the vector. C57BL/6-derived ES cells were electroporated with the IL-15 targeting vector and selected in G418 and ganciclovir as described 22. ES cell clones carrying a targeted IL-15 gene were identified by a combination of PCR and genomic Southern blot analyses and were injected into BALB/c blastocysts. Resulting male chimeras were bred to C57BL/6 females. The offspring were analyzed for germline transmission of the mutant IL-15 allele by PCR and genomic Southern blot analyses. Mice heterozygous for the IL-15 mutation (IL-15+/−) were intercrossed to generate IL-15–deficient (IL-15−/−) mice.
The IL-15−/− mice used throughout these studies were bred at Immunex and maintained on a C57BL/6 genetic background. All mice were housed under specific pathogen-free conditions and were used between 9 and 20 wk of age. Age- and sex-matched littermates (IL-15+/+ or IL-15+/−) or C57BL/6 mice (Taconic Farms, Inc.) were used as controls as indicated. Initial studies revealed no apparent difference between IL-15+/+ and IL-15+/− mice in cellularity of their secondary lymphoid tissues, phenotype of spleen or LN cells, or splenic NK cell responses.
Kennedy M.K., Glaccum M., Brown S.N., Butz E.A., Viney J.L., Embers M., Matsuki N., Charrier K., Sedger L., Willis C.R., Brasel K., Morrissey P.J., Stocking K., Schuh J.C., Joyce S, & Peschon J.J. (2000). Reversible Defects in Natural Killer and Memory Cd8 T Cell Lineages in Interleukin 15–Deficient Mice. The Journal of Experimental Medicine, 191(5), 771-780.
Publication 2000
Alleles Amino Acids antibiotic G 418 Blastocyst Cells Chimera Clone Cells Cloning Vectors Embryonic Stem Cells Exons Females Ganciclovir Genes Genetic Background Genome Germinal Center Germ Line Heterozygote Homologous Recombination Interleukin-15 Males Mice, Inbred C57BL Mus Mutation Natural Killer Cells Phenotype Southern Blotting Specific Pathogen Free Spleen Thymidine Kinase Transmission, Communicable Disease
4
Transgenic Mouse Model for Liver Cell Transplantation
Cited 17 times

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Publication 2011
Animals, Laboratory Biotransformation Cells Cell Transplantation Cloning Vectors Gene Expression Hepatocyte Homo sapiens Liver Liver Transplantations Mice, Inbred NOD Simplexvirus Strains Thymidine Kinase Transplantation Zygote
5
Aggrecan Knockin Mice Generation
Cited 15 times
The targeting vector plasmid for the Aggrecan knockin mice contains a 2.4 kb 5′ homologous arm, an IRES-CreERT2 cassette followed by a Pgk-neomycin (neo) cassette flanked with FRT sites, a 3.7 kb 3′ homologous arm, and a MC1-thymidine kinase cassette outside of the homology (Indra et al., 1999 (link); Mansour et al., 1988 (link)). The homologous arms were generated by PCR amplification using BAC template DNA from a clone isolated by a screening of a RPCI-22 129S6 Mouse BAC library (BAC PAC Resources). The nucleotide sequence of the PCR products were sequenced, and compared to sequences deposited in the mouse genome database using the BLAT algorithm (Kent et al., 2002 (link)). In the targeting vector, the neo cassette is in the reverse transcriptional orientation to the IRES-CreERT2 cassette. G4 ES cells (George et al., 2007 (link)) were electroporated with 25 micrograms of linearized targeting vector and placed under positive and negative selection according to standard procedures. Correctly targeted ES cell clones were screened by Southern blotting with flanking 5′ and 3′ probes. Chimeras generated by injection of C57BL/6 blastocysts were bred to C57BL/6 mice and all of the analysis was conducted in a mixed 129S6 × C57BL/6 genetic backround. Heterozygous Rosa26 reporter mice (Soriano, 1999b (link)) were bred to heterozygous aggrecan knockin mice (Agc1tm(IRES-creERT2)/+) and tissues from resulting progeny containing one copy of the R26R allele and the Aggrecan knockin allele were stained with X-gal (Gold Bio Technology). We used heterozygous Agc1tm(IRES-creERT2)/+ mice for all experiments, and the Frt-flanked Neo cassette was not removed.
Henry S.P., Jang C.W., Deng J.M., Zhang Z., Behringer R.R, & deCrombrugghe B. (2009). Generation of Aggrecan-CreERT2 knockin mice for Inducible Cre Activity in Adult Cartilage. Genesis (New York, N.Y. : 2000), 47(12), 805-814.
Publication 2009
5-bromo-4-chloro-3-indolyl beta-galactoside Aggrecans Alleles Arm, Upper Blastocyst Chimera Clone Cells Cloning Vectors DNA Library Embryonic Stem Cells Genome Gold Heterozygote Internal Ribosome Entry Sites Mice, Inbred C57BL Mus Neomycin Plasmids Reverse Transcription Thymidine Kinase Tissues

Most recents protocols related to «Thymidine Kinase»

1
Generation of In-frame Deletion Mutants
In-frame deletion mutants were generated using the suicide plasmid pEXG2 as described in Klein et al.34 (link) or its derivate pEXTK. In pEXTK, the sacB gene is replaced by a thymidine kinase gene. If pEXTK based mutator plasmids were used in the mutagenesis procedure, the positive selection to obtain a second crossover was performed by incubating merodiploidic clones for 3 h in 5 ml LB medium containing IPTG (1 mM). Subsequently, bacteria were positively selected by streaking bacteria on LB agar plates containing 200 µg/ml azidothymidine (Acros Organics) and 1 mM IPTG. Bacteria were then tested for loss of gentamicin sensitivity and mutants were verified by PCR as described previously34 (link). In brief, genomic DNA was isolated using the DNeasy Ultraclean Microbial Kit (Qiagen) and approximately 20 ng DNA was used to perform PCR using Mango Mix (Bioline), primers (Sigma-Aldrich) and water. Two primer pairs were used, namely (i) gene of interest (GOI) seqF and GOI seqR and (ii) GOI seqF and GOI insideR to distinguish between wildtype and mutant. The generated mutants and the primers used in this study are listed in Supplementary Data 1 or 2, respectively.
Eggers O., Renschler F.A., Michalek L.A., Wackler N., Walter E., Smollich F., Klein K., Sonnabend M.S., Egle V., Angelov A., Engesser C., Borisova M., Mayer C., Schütz M, & Bohn E. (2023). YgfB increases β-lactam resistance in Pseudomonas aeruginosa by counteracting AlpA-mediated ampDh3 expression. Communications Biology, 6, 254.
Publication 2023
Agar Bacteria Clone Cells Deletion Mutation Genes Genome Gentamicin Hypersensitivity Isopropyl Thiogalactoside Mangifera indica Mutagenesis Oligonucleotide Primers Plasmids Reading Frames Thymidine Kinase Zidovudine
2
Generation of Recombinant MVA Expressing SARS-CoV-2 RBD
For the generation of recombinant MVA expressing SARS-CoV-2 RBD wt and RBD M7 the shuttle vectors pMVA Trans TK-SARS-2 RBD wt and pMVA Trans TK SARS-2 RBD M7 were cloned. The MVA shuttle vectors were designed in a way that the antigens SARS-CoV-2 RBD wt and SARS-CoV-2 RBD M7 antigens can be inserted into the thymidine kinase (TK) locus J2R of the parental virus MVA CR19 TK-GFP under the transcriptional control of the early/late modified H5 promoter (mH5) via homologous recombination. The MVA shuttle vectors also include the reporter gene β-galactosidase (β-Gal) between the two left arm sequences of the TK locus for screening of recombinant MVAs. After several plaque purification rounds the reporter gene is lost by internal homologous recombination events resulting in a pure (reporter-free) recombinant MVA.
Carnell G.W., Billmeier M., Vishwanath S., Suau Sans M., Wein H., George C.L., Neckermann P., Del Rosario J.M., Sampson A.T., Einhauser S., Aguinam E.T., Ferrari M., Tonks P., Nadesalingam A., Schütz A., Huang C.Q., Wells D.A., Paloniemi M., Jordan I., Cantoni D., Peterhoff D., Asbach B., Sandig V., Temperton N., Kinsley R., Wagner R, & Heeney J.L. (2023). Glycan masking of a non-neutralising epitope enhances neutralising antibodies targeting the RBD of SARS-CoV-2 and its variants. Frontiers in Immunology, 14, 1118523.
Publication 2023
Antigens beta-Galactosidase Dental Plaque Genes, Reporter Homologous Recombination Parent SARS-CoV-2 Severe Acute Respiratory Syndrome Shuttle Vectors Thymidine Kinase thymidine kinase 2 Transcription, Genetic Virus
3
Generation of NF-κB Reporter Mice
The NF-κB reporter construct contained 4 repeats of NF-κB responsive element (5'-GGGAATTTCC-3'), minimal TA promoter (5'-TAGAGGGTATATAATGGAAGCTCGAATTCCAG-3'), Luciferase reporter gene, ubiquitin C (UBC) promoter, and the tdTomato fluorescence gene. Minimal TA promoter includes the TATA box from Herpes simplex virus thymidine kinase (HSV-tk) promoter. The firefly luciferase gene was amplified from NF-κB-luciferase plasmid (Clontech, CA, USA). To produce the targeting vector, the NF-κB reporter construct was cloned into the FseI site of Ai9 vector22 (link). The targeting vector was linearized with KpnI and electroporated into mouse embryonic stem cells (mES)23 (link), and 192 mES cell colonies that are tdTomato-positive and neomycin-resistance were picked. Chimeric males were bred with C57BL/6 females and the heterozygotes were backcrossed to C57BL/6 for at least 6 generations.
Park S.Y., Kim M.W., Kang J.H., Jung H.J., Hwang J.H., Yang S.J., Woo J.K., Jeon Y., Lee H., Yoon Y.S., Seong J.K, & Oh S.H. (2023). Novel NF-κB reporter mouse for the non-invasive monitoring of inflammatory diseases. Scientific Reports, 13, 3556.
Publication 2023
Chimera Cloning Vectors Females Fluorescence Genes Genes, Reporter Heterozygote Luciferases Luciferases, Firefly Males Mouse Embryonic Stem Cells Neomycin Plasmids RELA protein, human Simplexvirus TATA Box tdTomato Thymidine Kinase Ubiquitin C
4
NF-κB Signaling in Tibial Fracture Model
NF-κB-GFP reporter mice (Strain#: 027529) on a C57BL/6 background were purchased from the Jackson Laboratory (17 (link)), in which a transgenic construct contains an enhanced GFP-luciferase fusion gene under the control of tandem copies of a 36-base enhancer (containing two NF-κB binding sites) upstream of a herpes simplex virus minimal thymidine kinase promoter. Mice were housed in micro-isolator technique rodent rooms. We used 4-month-old male mice in the current study to exclude the effects of variations in levels of female sex hormones. Open tibial fractures were performed according to the standard operating procedure established in the URMC Center for Musculoskeletal Research (18 (link)). In brief, a 5 mm long incision was made in the skin on the anterior side of the tibia after anesthesia. A sterile 27 G needle was inserted into the marrow cavity of the tibia from the proximal end, temporarily withdrawn to facilitate midshaft transection of the tibia using a scalpel, and then reinserted to stabilize the fracture. The incision was closed with 5-0 nylon sutures. Mice received buprenorphine SR, 0.5 mg/kg to control pain. Fractures were confirmed by radiography using a Faxitron device (Hologic, Marlborough, MA). All animal procedures were conducted in accordance with approved guidelines of the University of Rochester Committee for Animal Resources (protocol number: 2001-121R).
Liu J., Lin X., McDavid A., Yang Y., Zhang H., Boyce B.F, & Xing L. (2023). Molecular signatures distinguish senescent cells from inflammatory cells in aged mouse callus stromal cells. Frontiers in Endocrinology, 14, 1090049.
Publication 2023
Anesthesia Animals Animals, Transgenic Binding Sites Buprenorphine Dental Caries Females Fracture, Bone Gene Fusion Gonadal Steroid Hormones Hormones Luciferases Males Marrow Medical Devices Mice, Laboratory Needles Nylons Pain RELA protein, human Rodent Simplexvirus Skin Sterility, Reproductive Strains Sutures Thymidine Kinase Tibia Tibial Fractures X-Rays, Diagnostic
5
Bacterial Genome Sequencing and Annotation
For the bacterial isolates, extracted DNA underwent library preparation and whole genome shotgun sequencing using the Illumina Miseq PE150 system at SeqCenter. The resulting bacterial genomes were assessed for their quality using FastQC version 0.11.9 and the resulting FastQC files were visualized using MultiQC version 1.12.60 ,61 (link) Sequences were then assembled and annotated using Bactopia version 2.2.1.62 (link) Genome quality was verified using CheckM (version 1.2.0).63 (link) Assembled genes underwent functional annotation, orthology assignments, and domain prediction using eggNOG-mapper v2.64 (link) All tools were used with default settings.
For the pyrimidine salvage gene pathway to be considered complete, all three of the following proteins needed to be present: thymidine kinase (K00857), thymidylate kinase (K00943), and nucleoside-diphosphate kinase (K00940).
For the de novo pyrimidine synthesis pathway to be considered complete, all eight of the following proteins (from KEGG Module 00051) needed to be present: dihydroorotate dehydrogenase (K00226 or K00254 or K17828), aspartate carbamoyltransferase catalytic subunit (K00609), aspartate carbamoyltransferase regulatory subunit (K00610), orotate phosphoribosyltransferase (K00762), dihydroorotase (K01465), orotidine-5’-phosphate decarboxylase (K01591), carbamoyl-phosphate synthase large subunit (K01955), and carbamoyl-phosphate synthase small subunit (K01956).
For the pyrimidine deoxyribonucleoside biosynthesis pathway to be considered complete, all six of the following proteins (from KEGG Module 00053) needed to be present: ribonucleoside-diphosphate reductase alpha chain (K00525), ribonucleoside-diphosphate reductase beta chain (K00526), thymidylate synthase (K00560), nucleoside-diphosphate kinase (K00940), thymidylate kinase (K00943), and dUTP pyrophosphatase (K01520).
Beauchemin E.T., Hunter C, & Maurice C.F. (2023). Actively replicating gut bacteria identified by 5-ethynyl-2’-deoxyuridine (EdU) click chemistry and cell sorting. Gut Microbes, 15(1), 2180317.
Publication 2023
Aspartate Carbamoyltransferase Bacteria Biosynthetic Pathways Carbamoyl-Phosphate Synthase (Ammonia) Catalytic Domain Deoxyribonucleosides Dihydroorotase Dihydroorotate Dehydrogenase DNA Library dUTP pyrophosphatase Genes Genome Genome, Bacterial Nucleoside-Diphosphate Kinase Orotidine-5'-Phosphate Decarboxylase Phosphoribosyltransferase, Orotate Proteins Protein Subunits Pyrimidines Ribonucleoside Diphosphate Reductase Thymidine Kinase thymidylate kinase Thymidylate Synthase

Top products related to «Thymidine Kinase»

1
Dual luciferase reporter assay system by Promega
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The Dual-Luciferase Reporter Assay System is a laboratory tool designed to measure and compare the activity of two different luciferase reporter genes simultaneously. The system provides a quantitative method for analyzing gene expression and regulation in transfected or transduced cells.
2
Lipofectamine 2000 by Thermo Fisher Scientific
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Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.
3
Prl tk by Promega
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The PRL-TK is a lab equipment product offered by Promega. It is a tool used for the detection and quantification of prolactin (PRL) levels. The core function of the PRL-TK is to provide researchers with a reliable method for analyzing PRL concentrations in various biological samples.
4
Dual luciferase assay kit by Promega
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The Dual Luciferase Assay Kit is a laboratory tool that measures the activities of two different luciferase reporter enzymes simultaneously within the same sample. The kit provides reagents and protocols to quantify the activities of firefly and Renilla luciferase reporters, allowing for normalization of experimental data.
5
Passive lysis buffer (plb) by Promega
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Passive lysis buffer is a solution used for the gentle lysis of cells to extract proteins or other biomolecules. It facilitates the release of cellular contents without denaturing the target analytes.
6
Dual glo luciferase assay system by Promega
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The Dual-Glo Luciferase Assay System is a reagent-based detection kit designed to quantify firefly and Renilla luciferase reporter gene activities in a single sample. The system provides a simple, sensitive, and reliable method for cell-based reporter gene analysis.
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Lipofectamine 3000 by Thermo Fisher Scientific
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Lipofectamine 3000 is a transfection reagent used for the efficient delivery of nucleic acids, such as plasmid DNA, siRNA, and mRNA, into a variety of mammalian cell types. It facilitates the entry of these molecules into the cells, enabling their expression or silencing.
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Dual luciferase assay system by Promega
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The Dual Luciferase Assay System is a laboratory tool designed to quantitatively measure the activity of two different luciferase reporter enzymes within the same sample. It provides a rapid and sensitive method for studying gene expression and regulation in a variety of experimental systems.
9
Dual luciferase reporter assay by Promega
Sourced in United States, Germany, China, United Kingdom, France, Switzerland, Japan
The Dual-Luciferase Reporter Assay is a laboratory equipment product that measures the activity of two different luciferase reporter enzymes simultaneously. It provides a method for analyzing gene expression, signal transduction pathways, and other biological processes in cells.
10
Fetal bovine serum (fbs) by Thermo Fisher Scientific
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.

More about "Thymidine Kinase"

Thymidine kinase (TK) is a crucial enzyme involved in DNA synthesis and repair, catalyzing the phosphorylation of thymidine to thymidine monophosphate, a key step in the pyrimidine nucleotide salvage pathway.
This enzyme plays a vital role in cell proliferation and is a marker for cellular growth and division, making it an important target for research and therapeutic development in areas such as cancer, viral infections, and gene therapy.
Researchers often utilize various reporter systems and assays to study TK activity, including the Dual-Luciferase Reporter Assay System, which allows for the simultaneous measurement of two different luciferase reporter enzymes, such as firefly luciferase (driven by the TK promoter) and Renilla luciferase (for normalization).
Lipofectamine 2000 and Lipofectamine 3000 are common transfection reagents used to introduce reporter plasmids, such as PRL-TK, into cells.
The Dual luciferase assay kit and Dual-Glo Luciferase Assay System provide reagents and protocols for the quantitative analysis of luciferase activity, while Passive lysis buffer is used to lyse cells and prepare samples for the assay.
By leveraging PubCompare.ai's AI-powered platform, researchers can optimize their TK studies by easily locating the best protocols, pre-prints, and patents from the literature, enhancing reproducibility and accuracy through intelligent analysis to identify the most effective methods and products.
This can help take your Thymidine kinase research to the next level, whether you're working in the context of cancer, viral infections, gene therapy, or other related fields.