|
Designing and Bioinformatic Analysis of Immunogenic Recombinant Chimeric Protein Containing rSEB and CTB for Vaccine
|
|
| Author:
|
FATEMEH MAGHSOOD AHMADI, ARASH MAHBOUBI, FARZANEH HOSSEINI, DAVOUD ESMAEILI, BAHAREH HAJIKHANI , SHAHRAM NAZARIAN
|
| Abstract:
|
Staphylococcal Enterotoxin B (SEB) is a bacterial antigen that is responsible for food poisoning in humans. Cholera toxin is the most important virulence factor in Vibrio cholera. The gene sequence of ctxB and seb were obtained from GenBank and Site-specific mutagenesis (L45R, Y89A, and Y94A, respectively) was performed in the seb gene sequence, Three-dimensional protein structure, modeling, secondary and tertiary structure, immunogenicity, and allergenic sites were predicted using online bioinformatics software. In addition to gene codons to increase the expression of the rSEB-CTXB protein in the E.coli BL21 DE3 host, different parameters affecting expression were also optimized. The thermodynamic analysis of mRNA structure was performed to investigate the stability at the transcriptional stage. The physicochemical parameters and the predicted protein structure and quality of the structures were predicted. Linear and conformational epitopes were also considered. The antigen tendencies and estimated sensitivity were performed using Algpred software. The chimeric protein is topologically close to native proteins and all domains had surface accessibility and showed the highest antigenicity index. The Codon Adaptation Index of chimer increased from 0.7 to 0.89. The protein structure showed that the predicted structure had good quality. The evaluation of silicon chimeric proteins showed that the model is suitable for the stability and solubility of mRNA. Constitutive and linear epitopes were observed in all three chimeric protein domains. This study showed that these results could have implications for the production of chimeric immunogen antigen against the toxicity of Staphylococcus aureus and Vibrio cholerae in animal models.The ctxB and Seb genes were examined for rare codons, and the genes were optimized, followed by bioinformatics. Protein expression was assessed by PAGE-SDS and Western blotting.The expression of recombinant protein from the synthetic gene led to the production of a protein with a size 51 kD.Conclusion: According to the results of bioinformatics studies and recombinant protein expression, it can be used as an immunogen for safety studies against Vibrio cholerae producing Cholera toxin and Staphylococcus aureus producing enterotoxin SEB.
|
| Keyword:
|
Chimeric, Gene ctxB, Insilco, Analysis rseb, Vaccine
|
| EOI:
|
-
|
| DOI:
|
https://doi.org/10.31838/ijpr/2021.13.01.822
|
| Download:
|
Request For Article
|
|
|