The use of a cell-free protein synthesis for obtaining bacterial diguanylatcyclase and two chimeric proteins
The novel promising trend of biotechnology is cell-free synthesis of proteins. Possibility of producing two chimeric proteins: adenosine deaminase of Escherichia coli fused with human annexin-A5, DNA-affine domain of bacteria Sulfolobus solfataricus fused with modified Taq-DNA-polymerase and bacterial enzyme diguanylatecyclase of Thermotoga maritima, by cell-free synthesis procedure as an alternative to conventional cultivation of microbial strains-producers in a bioreactor was studied in this research. Chimeric RNA polymerase of T7 bacteriophage, S30-cell extract of E. coli and multicopy plasmid pET42mut were engaged for protein synthesis by cell-free protein synthesis system. The first successful production of these proteins was demonstrated in CFPS system.