مقالات پذیرفته شده در سومین کنگره بین المللی زیست پزشکی
In silico design of multi-epitope vaccine against Pseudomonas aeruginosa; a bioinformatic approach
In silico design of multi-epitope vaccine against Pseudomonas aeruginosa; a bioinformatic approach
Maryam Gholami,1,*
1. Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
Introduction: Fusion protein technology is a cost-effective, high-speed strategy for protein expression. In most cases, immunogens are not yet able to induce strong Responses. The immune response focused on a specific immunodominant epitope is useful because it allows the strongest immune response against a certain pathogen to dominate, thus eliminating the pathogen fast and effectively. In addition, the epitope-focused immunogen capable to elicit convenient immune responses and efficiently boosts antibodies. Pseudomonas aeruginosa is an opportunistic pathogen affecting immunocompromised patients. It is known as one of the leading causes of nosocomial infections. A number of different vaccines and several monoclonal antibodies have been developed in the last decades for active and passive vaccination against the Gram-negative opportunistic pathogen Pseudomonas aeruginosa. It encodes a wide array of virulence factors and employs several strategies to evade immune detection. The alkaline protease (Apr) of Pseudomonas aeruginosa has been associated with bacterial virulence. In silico studies are a promising approach to design new chimeric fusion protein having the immunogenic potential of several antigens.
Methods: In the project to design a candidate vaccine from high antigenic regions of alkaline protease (A and B fragments;aprA ,aprB) as a recombinant fusion protein that contains two high-epitope-focused protein components that are linked by a linker fragment. At the first step primer was designed based on the secondary structure of the protein alkaline protease (outside the α-helix and β -sheet regions); For the step of purification and cleavage, designed reverse primer include His tag and enterokinase cleavage sites at C terminus. At the next step the nucleotide sequences were used to design a chimeric fusion gene which is constructed by aprA and aprB linked together by the RGRGGGGSGGGGGSSVEL fragment as a linker. The final step secondary and 3D structures and specificities of fusion protein were predicted by bioinformatics software.
Results: The results of this study showed that most regions of the chimeric fusion protein were found to have a high antigenic propensity and surface accessibility. Results showed that the designed fusion gene construction is suitable for chimeric fusion protein expression.
Conclusion: Finally, based on bioinformatic software analysis, designed chimeric fusion protein aprAB would be useful in animal model application and accounted candidate for multi-epitope vaccine development against Pseudomonas aeruginosa.