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- HAT_medium abstract "HAT Medium (hypoxanthine-aminopterin-thymidine medium) is a selection medium for mammalian cell culture, which relies on the combination of aminopterin, a drug that acts as a powerful folate metabolism inhibitor by inhibiting dihydrofolate reductase, with hypoxanthine (a purine derivative) and thymidine (a deoxynucleoside) which are intermediates in DNA synthesis. The trick is that aminopterin blocks DNA de novo synthesis, which is absolutely required for cell division to proceed, but hypoxanthine and thymidine provide cells with the raw material to evade the blockage (the \"salvage pathway\"), provided that they have the right enzymes, which means having functioning copies of the genes that encode them.The enzyme dihydrofolate reductase, which produces tetrahydrofolate (THF) by the reduction of dihydrofolate, is specifically blocked by aminopterin. THF, acting in association with specific proteins, can receive single carbon units that are then transferred to specific targets.One of the important targets for cellular reproduction is thymidylate synthase, which creates thymidine monophosphate (TMP) from deoxyuridine monophosphate (dUMP). By additional phosphorylation reactions, TMP can be used to make thymidine triphosphate (TTP), one of the four nucleotide precursors that are used by DNA polymerases to create DNA. Without the THF required to convert dUMP, there can be no TTP, and DNA synthesis cannot proceed, unless TMP can be produced from another source. The alternative source is the thymidine present in the HAT medium that can be absorbed by the cells and phosphorylated by thymidine kinase (TK) into TMP.The synthesis of IMP, (precursor to GMP and GTP, and to AMP and ATP) also requires THF, and also can be bypassed. In this case hypoxanthine-guanine phosphoribosyltransferase (HGPRT) reacts hypoxanthine absorbed from the medium with PRPP, liberating pyrophosphate, to produce IMP by a salvage pathway.Therefore, the use of HAT medium for cell culture is a form of artificial selection for cells containing working TK and HGPRT. Many useful refinements to the scheme are made possible by poisons that kill cells, but to which they are immune if they lack one of these genes. Thus, a cell lacking TK is resistant to bromodeoxyuridine (BrdU) and a cell lacking HGPRT is resistant to 6-thioguanine (6-TG) and 8-azaguanine. Thus, selection with one of the latter two drugs, followed by HAT medium, will yield revertant colonies.".
- HAT_medium thumbnail HAT_Selection.png?width=300.
- HAT_medium wikiPageID "3210816".
- HAT_medium wikiPageLength "4557".
- HAT_medium wikiPageOutDegree "54".
- HAT_medium wikiPageRevisionID "660990838".
- HAT_medium wikiPageWikiLink 8-Azaguanine.
- HAT_medium wikiPageWikiLink Aminopterin.
- HAT_medium wikiPageWikiLink Antibody.
- HAT_medium wikiPageWikiLink Antigen.
- HAT_medium wikiPageWikiLink B_cell.
- HAT_medium wikiPageWikiLink Biological_immortality.
- HAT_medium wikiPageWikiLink Bromodeoxyuridine.
- HAT_medium wikiPageWikiLink Category:Growth_media.
- HAT_medium wikiPageWikiLink Cell_division.
- HAT_medium wikiPageWikiLink César_Milstein.
- HAT_medium wikiPageWikiLink DNA.
- HAT_medium wikiPageWikiLink DNA_polymerase.
- HAT_medium wikiPageWikiLink DNA_synthesis.
- HAT_medium wikiPageWikiLink De_novo_synthesis.
- HAT_medium wikiPageWikiLink Deoxyuridine_monophosphate.
- HAT_medium wikiPageWikiLink Dihydrofolate_reductase.
- HAT_medium wikiPageWikiLink Enzyme.
- HAT_medium wikiPageWikiLink Epitope.
- HAT_medium wikiPageWikiLink File:HAT_Selection.png.
- HAT_medium wikiPageWikiLink Folic_acid.
- HAT_medium wikiPageWikiLink Gene.
- HAT_medium wikiPageWikiLink Georges_J._F._Köhler.
- HAT_medium wikiPageWikiLink Hybridoma_technology.
- HAT_medium wikiPageWikiLink Hypoxanthine.
- HAT_medium wikiPageWikiLink Hypoxanthine-guanine_phosphoribosyltransferase.
- HAT_medium wikiPageWikiLink Monoclonal_antibody.
- HAT_medium wikiPageWikiLink Multiple_myeloma.
- HAT_medium wikiPageWikiLink Niels_Kaj_Jerne.
- HAT_medium wikiPageWikiLink Nucleoside.
- HAT_medium wikiPageWikiLink Phosphoribosyl_pyrophosphate.
- HAT_medium wikiPageWikiLink Phosphorylation.
- HAT_medium wikiPageWikiLink Polyethylene_glycol.
- HAT_medium wikiPageWikiLink Precipitation_(chemistry).
- HAT_medium wikiPageWikiLink Protein.
- HAT_medium wikiPageWikiLink Purine.
- HAT_medium wikiPageWikiLink Pyrophosphate.
- HAT_medium wikiPageWikiLink Revertant.
- HAT_medium wikiPageWikiLink Selective_breeding.
- HAT_medium wikiPageWikiLink Sendai_virus.
- HAT_medium wikiPageWikiLink Tetrahydrofolic_acid.
- HAT_medium wikiPageWikiLink Thymidine.
- HAT_medium wikiPageWikiLink Thymidine_kinase.
- HAT_medium wikiPageWikiLink Thymidine_monophosphate.
- HAT_medium wikiPageWikiLink Thymidine_triphosphate.
- HAT_medium wikiPageWikiLink Thymidylate_synthase.
- HAT_medium wikiPageWikiLink Tioguanine.
- HAT_medium wikiPageWikiLinkText "HAT medium".
- HAT_medium wikiPageUsesTemplate Template:Other_uses.
- HAT_medium subject Category:Growth_media.
- HAT_medium hypernym Medium.
- HAT_medium comment "HAT Medium (hypoxanthine-aminopterin-thymidine medium) is a selection medium for mammalian cell culture, which relies on the combination of aminopterin, a drug that acts as a powerful folate metabolism inhibitor by inhibiting dihydrofolate reductase, with hypoxanthine (a purine derivative) and thymidine (a deoxynucleoside) which are intermediates in DNA synthesis.".
- HAT_medium label "HAT medium".
- HAT_medium sameAs Q3314146.
- HAT_medium sameAs Milieu_HAT.
- HAT_medium sameAs Mezzo_HAT.
- HAT_medium sameAs m.08zk3f.
- HAT_medium sameAs Q3314146.
- HAT_medium wasDerivedFrom HAT_medium?oldid=660990838.
- HAT_medium depiction HAT_Selection.png.
- HAT_medium isPrimaryTopicOf HAT_medium.