Optimization of genetic constructs for high-level expression of the darbepoetin gene in mammalian cells |
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Authors: | R. R. Shukurov K. Yu. Kazachenko D. G. Kozlov A. A. Nurbakov E. N. Sautkina R. A. Khamitov Yu. A. Seryogin |
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Affiliation: | 1. FARMAPARK Company, Moscow, 117246, Russia 2. State Research Institute of Genetics and Selection of Industrial Microorganisms, Moscow, 117545, Russia
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Abstract: | Genetic constructs were designed in order to optimize darbepoetin production in CHO cells. They are characterized by a higher level of structural optimization of the darbepoetin gene, a higher gene dose, and the selection of promoter elements that have never been used before for this purpose. A transient transfection of CHO cells by the obtained constructs was performed. It was shown that each of the variable factors in the constructs influenced darbepoetin gene expression. A construct containing a doubled dose of the darbepoetin synthetic gene with optimized codon composition under the control of the CMV-EF1α chimerical promoter was proved to be the most efficient. |
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