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Recovery of V79 cell clones with different phenotypes in aminopterin- or azaserine-containing media used for the selection of HGPRT revertants
Authors:R Randazzo  S Bonatti  G Luccarini  A Abbondandolo
Institution:1. Istituto di Genetica dell''Università, Palermo, Italy;2. Istituto di Mutagenesi e Differenziamento C.N.R., Pisa Italy
Abstract:Three 6-thioguanine (6TG)-resistant mutants were mutagen-treated and selected for clones capable of growing in 2 selective media: HAT medium, containing aminopterin (AP) and HAS medium, containing L-azaserine (AS). Both 6TG-sensitive, wild-type clones and 6TG-resistant mutants were found among colonies growing in HAT medium, while only 6TG-sensitive clones grew in HAS medium. Time for expression was required by 6TG-resistant but not by 6TG-sensitive clones, that were fully expressed immediately after treatment. All HAT-resistant, 6TG-resistant clones which were analyzed proved to be resistant to AP. These data were interpreted as follows: in HAT medium, both HGPRT+ revertants and double mutants (HGPRT?, AP-resistant) were selected, while only HGPRT+ revertants were selected in HAS medium. Not all 6TG-resistant mutants were able to produce both classes of HAT-resistant clones.
Keywords:AP  aminopterin  AS  EMS  ethyl methanesulfonate  HAS and HAT medium  see Materials and methods  HGPRT  hypoxanthine-guanine phosphoribosyl transferase (EC 2  4  2  8)  6TG  6-thioguanine
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