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Optimization of genetic constructs for high-level expression of the darbepoetin gene in mammalian cells
Authors:R. R. Shukurov  K. Yu. Kazachenko  D. G. Kozlov  A. A. Nurbakov  E. N. Sautkina  R. A. Khamitov  Yu. A. Seryogin
Affiliation:1. FARMAPARK Company, Moscow, 117246, Russia
2. State Research Institute of Genetics and Selection of Industrial Microorganisms, Moscow, 117545, Russia
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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