The importance of protein sequence control through Bioinformatics tools to achieve a large amount of desirable proteins in protein purification projects

Despite the fact that many protein expression systems have been developed today, but making a pure proteinremains a problematic duo to the presence of intensive cloning, expression and purification steps. Recentbioinformatic tools with In-silico analysis have allowed scientists to successfully identify immunogenicproteins.In our study, the In-silico analysis of the MAP۲۱۹۱ gene sequence was carried out using Bioinformaticstools, along with in vitro experiments. The antigenicity and hydrophobicity analysis of the Mce proteinencoded by this gene was investigated using CLC Genomics Workbench ۷.۵.۱. In-silico prediction of theprimary structure of Mce protein showed the presence of a highly hydrophobic loop in the N-terminal aminoacid sequence of residues ۱۰ to ۳۲. Then a new Mce protein encoded with the MAP۲۱۹۱ gene was amplifiedand subcloned into E. coli. We have attempted to express Mce protein in a different state with positiveexpression control.Western blot analysis of the Mce protein and the control protein only showed the presence of a controlprotein. Comparing the results of the prediction of the primary structure of the Mce protein with theexperimental results confirmed that the expression of this protein was influenced by its hydrophobic nature.Our data support the hypothesis that the presence of a hydrophobic structure, in particular during the firstinitialization of amino acids in the protein chain, may influence the protein expression system. Ourinformation provide valuable guidance for future researchers to scan the protein sequence for the hydrophobicregion in the starting stages of protein purification to have a sense of the expected final protein yield andmake better informed choices.

کلیدواژه ها:

Bioinformatic ، In-silico ، Hydrophobicity ، Protein purification