Application of Bioinformatics and Genetic Engineering for Designing Optimized Cloning and Overexpression of Neutrophil Activating Protein of Helicobacter pylori in Escherichia coli

Background and Objective: As the main virulence factor of Helicobacter pylori, HP-NAP has an important role in immunoprotection against this pathogen. This antigen is a strong candidate as a part of multi-component vaccine in the clinical trial against this bacterium. Due to NAP importance, it was used in this study as a template for optimization of heterologous genes with a low A-T content and low expression in E. coli. Materials and Methods: A synthetic single gene that could reach the highest level of expression in the host was designed by using bioinformatics tools. Results: A number of factors that influence gene expression level were changed for HP-NAP gene optimizing: the codon usage bias in E. coli was changed the G+C content was upgraded from ۳۸% to ۴۵% and the stem-loop structure was broken. These could result to prolong of the half-life of the mRNA and overexpression of recombinant of HP-NAP protein up to ۸۰۰ mg per liter. Conclusion: Applying of bioinformatics tools was appropriated to optimize of HP-NAP overexpression in E. coli. From our results, it appears that combination of In Silico and experimental approach is a logical approach for expression of heterologous genes in another host.

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Keywords: Helicobacter pylori , Bioinformatics , Optimization , Recombinant HP-NAP , Cloning , Expression.