Cotransfer and phenotypic stabilisation of syntenic and asyntenic mink genes into mouse cells by chromosome-mediated gene transfer.
By means of metaphase chromosomes, the genes for mink thymidine kinase (TK) and hypoxanthine-phosphoribosyltransferase (HPRT) were transferred to mutant mouse cells, LMTK-, A9 (HPRT-) and teratocarcinoma cells, PCC4-aza 1 (HPRT-). Eighteen colonies were isolated from LMTK- (series A), 9 from A9 (series B) and none from PCC4-aza 1. The transformed clones contained mink TK or HPRT. Analysis of syntenic markers in series B demonstrated that one clone contained mink glucose-6-phosphate dehydrogenase (G6PD) and the other alpha-galactosidase; in series A, nine clones contained mink galactokinase (GALK) and six mink aldolase C (ALDC). Analysis of 12 asyntenic markers located in ten mink chromosomes showed the presence of only aconitase-1 (ACON1) (the marker of mink chromosome 12) in three clones of series A. The clones lost mink ACON1 between the fifth to tenth passages. Cytogenetic analysis established the presence of a fragment of mink chromosome 8 in eight clones of series A, but not in series B. The clones of series A lost mink TK together with mink GALK and ALDC during back-selection; in B, back-selection retained mink G6PD. No stable TK+ phenotype was detected in clones with a visible fragment of mink chromosome 8. Stability analysis demonstrated that about half of the clones of series B have stable HPRT+ phenotype whereas only three clones of series A have stable TK+ phenotype. It is suggested that the recipient cells, LMTK- and A9, differ in their competence for genetic transformation and integration of foreign genes.
D000818:Animals; D002875:Chromosomes; D005796:Genes; D008040:Genetic Linkage; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D007621:Karyotyping; D007739:L Cells; D051379:Mice / Q000235:genetics; D008907:Mink / Q000235:genetics*; D010641:Phenotype; D013724:Teratoma; D013937:Thymidine Kinase / Q000235:genetics; D014170:Transformation, Genetic
D007041:Hypoxanthine Phosphoribosyltransferase; D013937:Thymidine Kinase