Optimization of genetic constructs for high-level expression of the darbepoetin gene in mammalian cells

R. R. Shukurov, K. Yu. Kazachenko, D. G. Kozlov, A. A. Nurbakov, E. N. Sautkina, R. A. Khamitov, Yu. A. Seryogin, 

Applied Biochemistry and Microbiology (Impact Factor: 0.66). 12/2014; 50(9). DOI: 10.1134/S0003683814090051

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.

12.01.2015, 2389 просмотров.