Metabolic burden in recombinant CHO cells: effect ofdhfr gene amplification andlacZ expression.

Cytotechnology
M B GuD S Kompala

Abstract

Foreign protein production levels in two recombinant Chinese hamster ovary (CHO) cell lines were compared in cells transfected with different expression vectors. One vector pNL1 contained the gene for neomycin resistance (neo ( r )) and thelacZ gene which codes for intracellular β-galactosidase, with both genes controlled by the constitutive simian virus (SV40) promoter. The other vector CDβG contained the amplifiabledhfr gene andlacZ gene, controlled by the constitutive SV40 and cytomegalovirus (CMV) promoters, respectively. Cell growth and β-galactosidase expression were compared quantitatively after cells were selected in different concentrations of the neomycin analog G418 and methotrexate, respectively. A 62% reduction in growth rate occurred in recombinant CHO cells in which thelacZ anddhfr genes were highly amplified and expressed. In contrast, the combined effects of the unamplifiedneo ( r ) gene andlacZ gene expression on the growth kinetics were small. Any metabolic burden caused bylacZ gene expression, which was evaluated separately from the effect ofneo ( r ) gene expression, must be negligible, as higher expression of β-galactosidase (1.5×10(-6) units/cell) occurred in unamplified cells compared to the cells in whi...Continue Reading

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Citations

Apr 25, 2012·Applied Microbiology and Biotechnology·Eugéne van RensburgJohann F Görgens
Nov 13, 2008·Cytotechnology·Christel FengeMichael Wood
Nov 4, 2008·Biotechnology and Bioengineering·Janet ChusainowMiranda G S Yap
Dec 7, 2016·Biotechnology and Bioengineering·Jong Youn Baik, Kelvin H Lee
Jul 7, 2017·Biotechnology and Applied Biochemistry·Wu ZouMohamed Al-Rubeai
Dec 21, 2017·Scientific Reports·Quang Vinh PhanManfred Gossen
Jan 24, 2019·Bioprocess and Biosystems Engineering·Kahina MellahiOlivier Henry
Feb 22, 2011·Biotechnology and Bioengineering·Britta KrampeMohamed Al-Rubeai
May 9, 2019·Biotechnology Letters·N JiménezZ P Gerdtzen

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