1st International and 10th National Iranian Conference on Bioinformatics
Home Accepted Papers
The importance of protein sequence control through Bioinformatics tools to achieve a large amount of desirable proteins in protein purification projects
Paper ID : 1232-ICB10
Authors:
Zahra Hemati *
Department of Pathobiology, School of Veterinary Medicine, ShahreKord University, ShahreKord, Iran.
Abstract:
Despite the fact that many protein expression systems have been developed today, but making a pure protein remains a problematic duo to the presence of intensive cloning, expression and purification steps. Recent bioinformatic tools with In-silico analysis have allowed scientists to successfully identify immunogenic proteins.
In our study, the In-silico analysis of the MAP2191 gene sequence was carried out using Bioinformatics tools, along with in vitro experiments. The antigenicity and hydrophobicity analysis of the Mce protein encoded by this gene was investigated using CLC Genomics Workbench 7.5.1. In-silico prediction of the primary structure of Mce protein showed the presence of a highly hydrophobic loop in the N-terminal amino acid sequence of residues 10 to 32. Then a new Mce protein encoded with the MAP2191 gene was amplified and subcloned into E. coli. We have attempted to express Mce protein in a different state with positive expression control.
Western blot analysis of the Mce protein and the control protein only showed the presence of a control protein. Comparing the results of the prediction of the primary structure of the Mce protein with the experimental 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 first initialization of amino acids in the protein chain, may influence the protein expression system. Our information provide valuable guidance for future researchers to scan the protein sequence for the hydrophobic region in the starting stages of protein purification to have a sense of the expected final protein yield and make better informed choices.
Keywords:
Bioinformatic, In-silico, Hydrophobicity, Protein purification
Status : Paper Accepted (Poster Presentation)