Parasexual crosses and hybrid selection with a near haploid variant of the epithelioma papulosum cyprini, cell line EPC: a tissue culture model for the analyses of dominance, recessiveness, and complementation of mutant phenotypes.
We describe techniques for insertional mutagenesis of tissue-cultured piscine cells in which we use transfection with G418 and hygromycin B resistance-conferring plasmids, cell matings by electrofusion, and positive selections of fusion hybrids by dual challenge with the antibiotics G418 and hygromycin B. These techniques are designed to facilitate genetic and molecular analyses of tissue-cultured cells. The experiments were conducted with EPC-1, a new variant of the carp epithelioma cyprini cell line. EPC, with a near haploid number of chromosomes, EPC-1 retains cell morphology and growth characteristics of EPC, including anchorage independence, but shows a higher degree of contact inhibition. The number of metaphase chromosomes of EPC-1 is 53, as opposed to 96 reported for EPC.
Li
Z F
ZF
Institute of Human Genetics, University of Minnesota Health Sciences Center, Minneapolis, 55455.
Murphy
T
T
Gao
D
D
Ellis
D
D
Sauerbier
A
A
Sauerbier
W
W
D000818:Animals; D002347:Carps; D002455:Cell Division; D002459:Cell Fusion; D002460:Cell Line; D002999:Clone Cells; D003001:Cloning, Molecular; D003260:Contact Inhibition; D003433:Crosses, Genetic; D004351:Drug Resistance; D005799:Genes, Dominant; D005808:Genes, Recessive; D005816:Genetic Complementation Test; D005839:Gentamicins / Q000494:pharmacology; D006921:Hygromycin B / Q000494:pharmacology; D007621:Karyotyping; D016254:Mutagenesis, Insertional; D010641:Phenotype; D010957:Plasmids; D014162:Transfection
D005839:Gentamicins; D006921:Hygromycin B; C010680:antibiotic G 418