Restricted integration into genomic DNA following transfection of Chinese-hamster ovary cells with N-acetylgalactosamine-4-sulphatase cDNA.
The effect of using different conditions of electroporation for introducing the gene coding for human N-acetylgalactosamine-4-sulphatase into Chinese-hamster ovary (CHO) cells has been studied in an effort to increase the yield of recombinant protein. In each case the range of enzyme activities in the medium of transfected cells proved to be similar, with genetic analysis of a higher producer revealing only one gene copy integrated. Hence, allowing for position effects, the number of gene copies stably integrated into the CHO genome of all transfectants presumably was low, potentially a single copy in each. This inability to allow high-copy-number integration may suggest a constraint on the levels of this protein which can be tolerated by the cell.
Webb
E
E
Research and Development Division, CSL Ltd., Parkville, Victoria, Australia.
D000818:Animals; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D018076:DNA, Complementary / Q000235:genetics* / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D018274:Electroporation; D004790:Enzyme Induction; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D008970:Molecular Weight; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013050:Spectrometry, Fluorescence; D015238:Sulfotransferases / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
D018076:DNA, Complementary; D011994:Recombinant Proteins; C498300:CHST9 protein, human; C092527:N-acetylgalactosamine-4-sulfotransferase; D015238:Sulfotransferases