[The distribution of 4 genes (alpha GAL, PGK-1, HPRT and G6PD) on the X chromosome of the American mink (Mustela vison)]. / Raspolozhenie chetyrekh genov (al'fa GAL, PGK-1, HPRT i G6PD) na X-khromosome amerikanskoi norki (Mustela vison).

The segregation of X-linked markers (alpha GAL, PGK-1, HPRT and G6PD) was analysed in hybrids between gamma ray-irradiated mink fibroblasts and Chinese hamster cells, or between mink cells and mouse hepatoma cells. Based on the segregation data and the data of cytogenetics analysis of a few hybrids, the order of the mink genes was deduced as alpha GAL--PGK-1--HPRT--G6PD--qter. This order differs from that reported for human and murine genes, in spite of the very obvious similarity between G-banding of the mink and human X chromosomes. Therefore, at least one reversion is responsible for the differences observed for the human and mink X chromosomes.

Subchromosomal localization and order of GLA, PGK1, HPRT, and G6PD loci on the X chromosome of the American mink (Mustela vison).

Segregation of the X-linked mink markers alpha-galactosidase (GLA), phosphoglycerate kinase-1 (PGK1), hypoxanthine phosphoribosyltransferase (HPRT), and glucose-6-phosphate dehydrogenase (G6PD) was analyzed in hybrids of gamma-irradiated mink fibroblasts and Chinese hamster cells and in hybrids of nonirradiated mink fibroblasts and mouse hepatoma cells. Based on this analysis, the order of the four genes is GLA-PGK1-HPRT-G6PD on the mink X chromosome. Cytogenetic analysis of five mink x Chinese hamster hybrid clones containing mink GLA, PGK1, and HPRT, but lacking G6PD, tentatively localized mink G6PD to Xq15.22----qter and also confirmed the gene order as GLA-PGK1-HPRT-G6PD-qter. Comparison of this order with its counterpart in man and the mouse, as well as an analysis of the G-band patterns of their X chromosomes, demonstrated putative similarities between mink and man and differences in the mouse. These differences may be due to a different rate of X-chromosomal rearrangement in mammalian evolution.

Peptidases A, B, C, D and S in the American mink: polymorphism and chromosome localization.

An electrophoretic analysis of peptidases was carried out in a population of American mink. Based on substrate and tissue specificities, as well as subunit composition, homologies were established between mink peptidases A, B, C, D and S and human peptidases. Polymorphism for peptidases B and D was demonstrated for minks of three coat colour types. Breeding data indicated that the peptidase variations are under the control of allele pairs at distinct autosomal loci designated as PEPB and PEPD, respectively. Using a panel of American mink-Chinese hamster hybrid clones, the gene for PEPB was assigned to mink chromosome 9.

Transfer of mink genes into mouse cells by means of isolated lipid-encapsulated nuclei.

A method for gene transfer by means of interphase nuclei encapsulated within lipid membranes was developed. The method was based on passage of interphase nuclei through a layer of organic solvents containing phospholipids. Evidence was obtained indicating that the nuclei become surrounded by a protective phospholipid membrane: measurements of bound labelled or non-labelled phospholipids; decrease in the permeability of lipid-encapsulated nuclei for high molecular compounds; visualization by direct electron microscopy. Lipid-encapsulated nuclei of mink fibroblasts were used for transformation of mutant mouse LMTK- cells (deficient for thymidine kinase). The frequency of occurrence of HAT-resistant colonies/recipient cell was 1.9 X 10(-5). Biochemical analysis of 14 independent clones demonstrated that they all contained TK1 of mink origin. Analysis of 15 other biochemical markers located on 12 of the mink chromosomes revealed the activities of mink galactokinase (a syntenic marker) in 5 transformed clones, and that of mink aconitase-1 (the marker of mink chromosome 12) in 1 clone. No cytogenetically visible donor chromosomes were identified in the transformed clones. Nine transformed clones were tested for the stability of the TK+ phenotype; of these, the phenotype was expressed stably in 3 and unstably in 6. The method suggested is similar to the gene transfer procedure using total DNA. Its advantage is in ensuring efficient gene transfer and donor DNA integrity.

Regional assignments of eight genes on chromosome 2 in the American mink.

Chromosomal rearrangements involving mink chromosome 2 in mink-Chinese hamster and mink-mouse hepatoma somatic hybrids were identified. By means of these rearrangements, we assigned the genes for HK1, GOT1, and PP to 2pter----p22, those for PGD, PGM1 and ENO1 to 2q24.4----qter, and that for NP and ADK to 2pter----p11.1.

Regional assignment of the genes for TK1, GALK, ALDC, and ESD on chromosome 8 in the American mink by chromosome-mediated gene transfer.

A panel of clones of mink-Chinese hamster somatic cell hybrids was analysed to obtain data for assigning the genes for thymidine kinase-1 (TK1), galactokinase (GALK), subunit C of aldolase (ALDC), and esterase D (ESD) to specific mink chromosomes. The results demonstrate that the genes for TK1, GALK, ALDC and ESD are syntenic and located on mink chromosome 8. Prometaphase analysis of transformed mouse cells obtained by transfer of mink genes by means of metaphase chromosomes demonstrated the presence of mink chromosome 8 fragments of different sizes in some of the independent transformants. Segregation analysis of these fragments and mink TK1, GALK, ALDC and ESD allowed us to assign the genes for TK1 and GALK to 8p24, ALDC to pter-8p25, and ESD to 8q24-8qter.

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.

Chromosome localization of the genes for ENO1, HK1, ADK, ACP2, MPI, ITPA, ACON1 and α-GAL in the American mink (Mustela vison).

Twenty-eight American mink × Chinese hamster somatic cell hybrids were analysed for the expression of mink enzymes and the segregation of mink chromosomes. The results demonstrated that the gene for enolase-1 is located on the long arm of mink chromosome 2, and those for hexokinase-1 and adenosine kinase, on its short arm. Segregation analysis of mink chromosomes and mink acid phosphatase-2, mannose phosphate isomerase, inosine triphosphatase and aconitase-1 provided data allowing us to assign the genes for these markers to mink chromosomes 7, 10, 11 and 12, respectively. The expression of mink α-galactosidase was highly coincidental with mink × chromosome as well as with its markers: hypoxanthine-phosphoribosyltransferase, glucose-6-phosphate dehydrogenase and phosphoglycerate kinase-1. This result confirms the assignment of the gene for α-galactosidase to the mink × chromosome.

Chromosome localization of the genes for isocitrate dehydrogenase-1, isocitrate dehydrogenase-2, glutathione reductase, and phosphoglycerate kinase-1 in the American mink (Mustela vison).

Twenty-eight hybrid clones with different mink chromosomes were derived from the fusion of Chinese hamster and American mink (mustela vison) cells. This set of clones made it possible to assign the mink genes for isocitrate dehydrogenase-1 (soluble) to chromosome 4, for isocitrate dehydrogenase-2 (mitochrondrial) to chromosome 10, for glutathione reductase to chromosome 6, and for phosphoglycerate kinase-1 to the X chromosome.

Chromosome localization of the loci GOT1, PP, NP, SOD1, PEPA and PEPC in the American mink (Mustela vison).

Twenty eight American mink × Chinese hamster somatic cell hybrids were analysed for the expression of mink enzymes and chromosome segregation. This analysis made it possible to assign the genes for glutamate-oxaloacetate transaminase-1 (soluble) (EC 2.6.1.1), inorganic pyrophosphatase (EC 3.6.1.1), purine nucleoside phosphorylase (EC 2.4.2.1) to mink chromosome 2, superoxide dismutase-1 (soluble) (EC 1.11.1.1) to chromosome 5, peptidase A (EC 3.4.11 or 3.4.13) to chromosome 4, and peptidase C (EC 3.4.11 or 3.4.13) to chromosome 13. It is suggested that the synthenic gene group GOT1-PP-NP is located on the short arm of mink chromosome 2.

Chromosome localization of three syntenic gene pairs in the American mink (Mustela vison).

Twenty eight American mink X Chinese hamster somatic cell hybrids were analysed for the expression of mink enzymes and for mink chromosomes. The results of this analysis made it possible to assign the genes for phosphoglucomutase-1 and phosphogluconate dehydrogenase to chromosome 2, those for lactate dehydrogenase A and glucose phosphate isomerase to chromosome 7, and those for lactate dehydrogenase B and triosephosphate isomerase to chromosome 9.

[Isolation and characteristics of somatic cell hybrids of the Chinese hamster and American mink]. / Poluchenie i kharakteristika gibridov somaticheskikh kletok kitaiskogo khomiachka i amerikanskoi norki.

The paper deals with obtaining somatic cell hybrids of Chinese hamster and mink by means of inactivated Sendy virus. 39 hybrid clones segregating mink chromosomes were formed by fusing Chinese hamster cells deficient in hypoxanthine phosphoribosyliransferase with normal cells of mink. Enzyme analyses of these hybrid clones revealed that in mink genes coding lactate dehydrogenase-A, lactate dehydrogenase-B, malate dehydrogenase-NAD (soluble), 6-phosphogluconate dehydrogenase, glucose-6-phosphate dehydrogenase are not syntenic. A possibility of successful utilization of these somatic cell hybrids for mapping mink genes is shown.

Chinese hamster x American mink somatic cell hybrids: characterization of a clone panel and assignment of the mink genes for malate dehydrogenase, NADP-1 and malate dehydrogenase, NAD-1.

Chinese hamster x American mink somatic cell hybrids were obtained and examined for chromosome content and expression of mink malate dehydrogenase, NADP (MOD-1; EC 1.1.1.40), malate dehydrogenase, NAD (MOR-1; EC 1.1.1.37), glucose-6-phosphate dehydrogenase (G6PD; EC 1.1.1.49) and hypoxanthine phosphoribosyltransferase (HPRT; EC 2.4.2.8). All the hybrid clones examined were found to segregate mink chromosomes. A clone panel containing 25 clones was set up. The possibilities and limitations of this panel for mink gene mapping are analysed. Using this panel, it is feasible to rapidly map genes located on chromosomes 1-13 and to provisionally assign genes located on chromosome 14 and the X. Based on the data obtained, the genes for MOD-1 and MOR-1 were firmly assigned to mink chromosomes 1 and 11, respectively, and the genes for G6PD and HPRT were provisionally assigned to the X.