Reappearance of murine hypoxanthine guanine phosphoribosyl transferase activity in mouse A9 cells after attempted hybridisation with human cell lines
| Autor: | I.P. Gormley, H. Harris, B. Watson, S. E. Gardiner, H. J. Evans |
|---|---|
| Rok vydání: | 1972 |
| Předmět: |
Guanine
Reversion Glucosephosphate Dehydrogenase Hybrid Cells Biology Lymphocyte Activation Cell Line Mice chemistry.chemical_compound medicine Animals Humans Lymphocytes Pentosyltransferases Fibroblast Lung Cells Cultured Hypoxanthine Phosphoglycerate kinase Sex Chromosomes Lymphoblast Cell Biology Fibroblasts Embryo Mammalian Molecular biology Aminopterin Leukemia Lymphoid Phosphoglycerate Kinase medicine.anatomical_structure Genes chemistry Biochemistry Hypoxanthine-guanine phosphoribosyltransferase Cell culture Hypoxanthines Karyotyping Hybridization Genetic Thymidine |
| Zdroj: | Experimental Cell Research. 75:401-409 |
| ISSN: | 0014-4827 |
| DOI: | 10.1016/0014-4827(72)90446-6 |
| Popis: | A method for the electrophoresis of hypoxanthine guanine phosphoribosyl tranferase (HGPRT) on starch gels is described which can distinguish human and murine HGPRT and resolve each into a number of bands. Five putative hybrid lines, made between HGPRT-deficient (HGPRT − ) mouse fibroblasts (A9 cells) and HGPRT + human cells (lymphocytes, fibroblast and lymphoblast cell lines), and which were able to grow in the presence of HAT, were tested for their HGPRT type. The expected human HGPRT was seen in only two of the five lines and in one of these murine HGPRT was also present. The line with the mixture of HGPRT types lost the human enzyme on further subculture. The remaining three lines had only murine HGPRT. The reversion of HGPRT- to HGPRT + in A9 cells is an extremely rare event and the presence of murine HGPRT in four of the five cell lines studied, and in none of the control A9 cultures, indicates a very high rate of reversion (derepression) in fusion experiments with human cells. Where human HGPRT was detected, both human and murine glucose-6-phosphate dehydrogenase, together with a hybrid band, was seen, and also a mixture of human and murine phosphoglycerate kinase with no hybrid band. Human X chromosomes were cytologically demonstrable where the three human enzymes were present and in later subcultures of the line with no murine HGPRT, human PGK became lost whereas the two other human X-linked enzymes were consistently found. |
| Databáze: | OpenAIRE |
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