caa a22 4500 465820344 CHVBK 20180323112134.0 cr unu---uuuuu 170327e19901101xx s 000 0 eng 10.1007/BF01233092 doi (NATIONALLICENCE)springer-10.1007/BF01233092 Genetic analysis of a vital mammalian housekeeping locus using CHO cells that express a transfected mutant allele [Elektronische Daten] [J. Diaz, D. Rhoads, D. Roufa] We describe a novel approach for the isolation of null mutations in a vital Chinese hamster ovary (CHO) cell housekeeping gene. Our experimental strategy required introduction of an expressible DNA clone encoding a recessive emetine-resistance allele of ribosomal protein S14 into wild-type CHO cells. Transgene heterozygote (TGH) cell lines, which harbor multiple emetine-resistance S14 transgenes, survive mutations that inactivate the CHO RPS14 locus by virtue of the transgenes' biological function. Null mutations inRPS14 yield TGH clones that display the transgene's drug-resistance phenotype. A large collection of emetine-resistant clones was isolated from one TGH cell line and shown to consist of three types of S14 mutations: (1) nonsense null mutations in theRPS14 protein coding sequence; (2) missense null mutations that affect S14 amino acid residues that have been conserved stringently during eukaryotic evolution, and (3) a recurrent missense mutation that results in a new, functionalRPS14 emetine-resistance allele. Plenum Publishing Corporation, 1990 Diaz J. Division of Biology, Kansas State University, 66506, Manhattan, Kansas aut Rhoads D. Division of Biology, Kansas State University, 66506, Manhattan, Kansas aut Roufa D. Division of Biology, Kansas State University, 66506, Manhattan, Kansas aut Somatic Cell and Molecular Genetics Kluwer Academic Publishers-Plenum Publishers 16/6(1990-11-01), 517-528 0740-7750 16:6<517 1990 16 11188 https://doi.org/10.1007/BF01233092 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF01233092 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Diaz J. Division of Biology, Kansas State University, 66506, Manhattan, Kansas aut NATIONALLICENCE 700 1- Rhoads D. Division of Biology, Kansas State University, 66506, Manhattan, Kansas aut NATIONALLICENCE 700 1- Roufa D. Division of Biology, Kansas State University, 66506, Manhattan, Kansas aut NATIONALLICENCE 773 0- Somatic Cell and Molecular Genetics Kluwer Academic Publishers-Plenum Publishers 16/6(1990-11-01), 517-528 0740-7750 16:6<517 1990 16 11188 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer