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• W1520299277 abstract "The use of thymidine kinase as a selective marker in transfection experiments is established using the thymidine kinase, tk, gene of herpes simplex virus (HSV-tk). The application of tk as a negative marker changes dramatically when substrate analogs are developed that HSV-tk recognizes but for which the mammalian protein has no apparent affinity. The negative selection with acyclovir and GANC, ganciclovir {GANC-2-amino-9- [2-hydroxy-1- (hydroxymethyl)ethoxymethyl]-9H-purine-6(1H)-on}, is reported to be faster than that using FIAU [1-(2-deoxy-2-fluoro-β-o- rabinofuranosyl)-5-iodouracil], acyclovir{9-[(2-hydroxyethoxy)methyl]guanine}, and in this chapter only results with GANC is given. The use of tk as positive marker requires functional cassettes that can be selected positively in one phase of the experiment and negatively in another. The optimal solution would be the use of one single gene for such a task. An obvious next step is the combination of HSV-tk with other genes by gene fusion. A problem with these new fusion genes is that many cell lines already have a resistance to neomycin or hygromycin. Not all cells are readily selectable with these drugs. In these cases, another positive selectable gene should be fused to HSV-tk. There are two possible orientations, 5´ and 3´, to the other gene. Full length HSV-tk can be used, complete from the first codon to the last. No amino acid linker (spacer) between HSV-tk and the second part of the protein seems to be necessary for the negative selection to function, at least in the cases tested so far. The solution is to perform the actual fusion by polymerase chain reaction (PCR)." @default.
• W1520299277 created "2016-06-24" @default.
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• W1520299277 date "2000-01-01" @default.
• W1520299277 modified "2023-10-16" @default.
• W1520299277 title "[9] Use of fusions to thymidine kinase" @default.
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• W1520299277 doi "https://doi.org/10.1016/s0076-6879(00)26051-5" @default.
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