Genetics of Somatic Mammalian Cells: Linkage Studies with Human–Chinese Hamster Cell Hybrids

Nature ◽  
1970 ◽  
Vol 228 (5269) ◽  
pp. 329-332 ◽  
Author(s):  
FA-TEN KÁO ◽  
THEODORE T. PUCK
1981 ◽  
Vol 1 (4) ◽  
pp. 336-346
Author(s):  
C E Campbell ◽  
R G Worton

Somatic cell hybrids heterozygous at the emetine resistance locus (emtr/emt+) or the chromate resistance locus (chrr/chr+) are known to segregate the recessive drug resistance phenotype at high frequency. We have examined mechanisms of segregation in Chinese hamster cell hybrids heterozygous at these two loci, both of which map to the long arm of Chinese hamster chromosome 2. To follow the fate of chromosomal arms through the segregation process, our hybrids were also heterozygous at the mtx (methotrexate resistance) locus on the short arm of chromosome 2 and carried cytogenetically marked chromosomes with either a short-arm deletion (2p-) or a long-arm addition (2q+). Karyotype and phenotype analysis of emetine- or chromate-resistant segregants from such hybrids allowed us to distinguish four potential segregation mechanisms: (i) loss of the emt+- or chr+-bearing chromosome; (ii) mitotic recombination between the centromere and the emt or chr loci, giving rise to homozygous resistant segregants; (iii) inactivation of the emt+ or chr+ alleles; and (iv) loss of the emt+- or chr+-bearing chromosome with duplication of the homologous chromosome carrying the emtr or chrr allele. Of 48 independent segregants examined, only 9 (20%) arose by simple chromosome loss. Two segregants (4%) were consistent with a gene inactivation mechanism, but because of their rarity, other mechanisms such as mutation or submicroscopic deletion could not be excluded. Twenty-one segregants (44%) arose by either mitotic recombination or chromosome loss and duplication; the two mechanisms were not distinguishable in that experiment. Finally, in hybrids allowing these two mechanisms to be distinguished, 15 segregants (31%) arose by chromosome loss and duplication, and none arose by mitotic recombination.


1980 ◽  
Vol 102 (2) ◽  
pp. 209-216 ◽  
Author(s):  
Jennifer A. Marshall Graves ◽  
Kenneth W. Koschel

Genetics ◽  
1973 ◽  
Vol 75 (3) ◽  
pp. 515-530
Author(s):  
D J Roufa ◽  
B N Sadow ◽  
C T Caskey

ABSTRACT In order to obtain a large collection of Chinese hamster cell clones defective in thymidine kinase (TK-), BrdUr selection experiments have been performed on wild-type and revertant TK+ cell lines. No clones (< 10-9) were obtained from the wild-type TK+ cell line by single-step selection. In contrast, revertant TK+ clones readily gave rise to stable TK- derivatives (1 - 2 × 10-4). Both wild-type and revertant TK+ clones spontaneously yielded 8-AGr colonies with the same frequency (1 - 5 × 10-6), suggesting that the differences between wild-type and revertant cell lines specifically affected selection of the TK- phenotype. The increased frequency of TK- clones reflects perhaps the number (ploidy) or character of the autosomal TK loci in TK+ revertants, or perhaps the mechanisms which regulate expression of the TK genes. Several mutagens, EMS, MNNG and UV, stimulated the TK+ revertants' frequency of TK- subclones only slightly (< 3-fold). Biochemical and genetic data indicated that the TK- clones derived from one revertant are phenotypically different. The phenotypes displayed by these cell lines are stable and do not depend upon the continued presence of the selective agent.


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