Abstract
The maintenance of high plasmid copy-number and the over-expression of foreign proteins in recombinant bacteria and yeasts is known to result in a metabolic load, which adversely affects the growth rate. Reports suggest that recombinant mammalian systems are similarly affected, however in comparison to bacterial systems, relatively little information exists. It was the aim of this study to test the effects of recombinant gene expression on the growth and metabolism of two industrially important mammalian cell lines. BHK 570 and CHO-K1 cells were stably transfected with the human gastric inhibitory peptide (h-GIP) and glucagon receptor respectively. Selection was by way of the neomycin resistance (neo’) gene using G418 as the selective agent. On removal of G418, production of both receptors was stable for the course of the experiments. The growth and metabolism of both cell lines was affected by the presence of G418 in a manner indicative of an increased metabolic load, caused by over-expression of the neo’ protein. The two cell lines differed in their response to the metabolic load, suggesting a cell-line or clone dependent response. Growth under the increased metabolic load could be modulated by serum, insulin and glutamine addition to the growth medium. Implications for the use of G418 are discussed.
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Yallop, C.A., Svendsen, I. (2001). The Effects of Recombinant Protein Expression on the Growth and Metabolism of Mammalian Cells. In: Merten, OW., et al. Recombinant Protein Production with Prokaryotic and Eukaryotic Cells. A Comparative View on Host Physiology. Springer, Dordrecht. https://doi.org/10.1007/978-94-015-9749-4_3
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DOI: https://doi.org/10.1007/978-94-015-9749-4_3
Publisher Name: Springer, Dordrecht
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