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1- Department of Animal Science, Faculty of Agriculture, Lorestan University, Khorramabad, Iran , forouharmehr.a@lu.ac.ir
2- Department of Pathobiology, Faculty of Veterinary Medicine, Lorestan University, Khorramabad, Iran
Abstract:   (142 Views)
Background: The heparin-binding hemagglutinin protein, which belongs to Mycobacterium tuberculosis is known as a molecular adjuvant. Hence, the expression of this protein in the prokaryotic system is important.
Methods: To predict an appropriate signal peptide for expression of the heparin-binding hemagglutinin protein, 50 signal peptides were selected from the Signal Peptide database. Then, the most important parameters of signal peptides including, signal peptide probability, different regions of signal peptides, physicochemical features, signal peptide sorting and sub-cellular location were completely investigated by reliable tools. After identification of the best predicted signal peptide, it was linked to the heparin-binding hemagglutinin protein, and its secondary structure, tertiary structure and in silico cloning in pET21a (+) were assessed. 
Results: The results of different evaluations confirmed that only 13 signal peptides passed all features including, clearance of N, H, and C regions, D-score > 0.7, instability index>40, and periplasmic localization.  Finally, based on D-scores, among these 13 signal peptides, asr (acid shock RNA) peptide with D-score =90 was selected as the best predicted signal peptide to apply. Moreover, the results showed that the secondary structure of the adjuvant linked to asr peptide contained 88.18% alpha helix and 9.5% random coil. Also, the results of in silico cloning revealed that nucleotide sequence of the adjuvant linked to asr peptide were successfully cloned between BamHI and XhoI enzymes in the multiple cloning site of pET21a (+).
Conclusion: The results of this study confirmed that asr peptide can be used as an appropriate signal peptide for expression of the heparin-binding hemagglutinin protein in the prokaryotic system
     
Type of Study: Research | Subject: Biotechnology

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