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Cotransfer of syntenic human genes into mouse cells using isolated metaphase chromosomes or cellular DNA

Summary

Chromosome-mediated gene transfer (CMGT) of the human genes for hypoxanthine phosphoribosyl transferase (HPRT) and cytosol thymidine kinase (TK1) into HPRT deficient mouse A9 cells or TK deficient Swiss mouse 3T3TK cells was found to occur at frequencies at least one order of magnitude higher than DNA-mediated gene transfer (DMGT). The frequency of CMGT into 3T3TK cells was reduced by more than an order of magnitude by a posttreatment of the recipient cells with dimethyl sulphoxide (DMSO). After CMGT, expression of the non-selected genes coding for galactokinase (GALK) and acid alpha-glucosidase (GAA), both syntenic with TK1, was observed in a number of transformants. From the pattern of cotransfer, a tentative gene ordering of CENTROMERE-GALK-TK1-GAA on human chromosome 17 was deduced. Chromosome-mediated cotransfer of X-linked human phosphoglycerate kinase (PGK) with HPRT was observed in two out of 33 A9 transformants analysed. DNA-mediated cotransfer of a syntenic gene was only observed for GALK, cotransferred with TK1 in two out of 18 TK+ transformants of mouse LTK cells. Therefore, with murine cells as recipients of human donor genetic material, CMGT results in a higher frequency of transfer and a higher incidence of cotransfer of syntenic genes than DMGT using cellular DNA in the same cell system.

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Correspondence to A. J. J. Reuser.

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de Jonge, A.J.R., de Smit, S., Kroos, M.A. et al. Cotransfer of syntenic human genes into mouse cells using isolated metaphase chromosomes or cellular DNA. Hum Genet 69, 32–38 (1985). https://doi.org/10.1007/BF00295526

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Keywords

  • Thymidine Kinase
  • Dimethyl Sulphoxide
  • Phosphoglycerate Kinase
  • Phosphoribosyl
  • Phosphoribosyl Transferase