Cloning and Protein Production of the Antigenic FliC Gene of Salmonella enterica serovar typhimurium ATCC 14028 Strain

Proteins are large and complex biomolecules that perform a wide variety of biological functions in living organisms. These biomolecules play a critical role in many fields, including medicine, industry, food, the environment, and scientific research. Producing these proteins in their natural environment is both inefficient and extremely difficult. Using a molecular technique called recombinant DNA technology, proteins can be easily produced in appropriate quantities in an organism other than their natural source. The primary objective of this study was to clone the FliC gene from Salmonella enterica serovar typhimurium ATCC 14028 into the pET-SUMO vector for the first time and produce recombinant protein. For this purpose, the FliC gene obtained from S. typhimurium ATCC 14028 was inserted into the pET-SUMO vector, and recombinant protein production was carried out in Escherichia coli (E. coli) BL21(DE3) cells. The results showed that the culture induced with 1 mM IPTG provided the highest protein yield. The produced protein, approximately 70 kDa in size, was confirmed and purified using SDS-PAGE and nickel (Ni+2) affinity chromatography. In this study, the S. typhimurium FliC gene was successfully transferred to the pET-SUMO vector using the A-T cloning method for the first time, enabling the production of the targeted recombinant FliC protein.