KEY POINTS TO OBTAINING ARTIFICIAL RECOMBINANT PROTEINS IN PROKARYOTIC SYSTEMS

Abstract

Developed in the 1970s, recombinant DNA technology transformed biotechnology by enabling the production of recombinant proteins through molecular cloning. This technique has enabled the development of more effective diagnostics, drugs, and vaccines. However, the production of recombinant proteins, especially artificial ones, can be a challenging task. Researchers and technicians must consider cloning, production, and purification steps for satisfactory protein production and yield. The selection of vectors and expression systems, codon optimization, epitope prediction, and biochemical characteristics of the protein are some of the points to be considered during recombinant protein construction, especially for artificial ones. This article describes the main processes to obtain artificial recombinant proteins to help researchers and technicians with protein construction tasks. It compiles essential key points for a successful construction and expression. With a special focus on the prokaryotic system of Escherichia coli, we suggest software, in silico analyses, bench techniques, and reviews that range from the construction of the amino acid sequence to purification in satisfactory yield.