IN VIVO PRODUCTION OF INSULIN-LIKE GROWTH FACTOR CODED BY PLASMID DNA

Abstract

The aim was to explore the expression of recombinant insulin-like growth factor (IGF-1) after intramuscular electroporation of mice with plasmid DNA coding this protein. For this purpose synthetic gene coding IGF-1 was introduced into plasmid pcDNA3, giving pcDNA3igf-1 and Escherichia coli strain E.coli DH10 (pcDNA3igf-1) , the producer of this plasmid , was created. Highly purified plasmid pcDNA3igf-1 was used for intramuscular electroporation with the aid of the electric device designed at the laboratory to generate electric pulses. IGF-1 concentrations in sera were determined by ELISA. Testing several modes of electric pulses had shown that the most effective mode for IGF-1 expression in Balb/c mice in vivo was single pulse of square profile with the amplitude of 160 V and 2 0 milliseconds duration. After intramuscular injection of 5 0 micrograms of pcDNA3igf-1, followed by immediate electric pulse the maximal concentration of IGF-1 in serum increased to 324 ng/ml exceeding the control level 3,44 times and high level of IGF-1 was observed during 7 days after electroporation.

Keywords

insulin-like growth factor-1 (IGF-1) / plasmid / expression in vivo / electroporation

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RIGHTS & PERMISSIONS

Dukhovlinov I.V., Bogomolova E.G., Dobrovolskaya O.V., Fedorova E.A., Klaus A.M., Ishuk C.A., Klimov N.A., Simbirtsev A.S.