Bioinformatic Design and Construction of a Chimeric Gene Comprising the stxB, ctxB and blf1 Genes

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MOSHIRI ZAHRA; HADDAD RAHEEM; HONARI HOSSEIN

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Journal: JOURNAL OF NEYSHABUR UNIVERSITY OF MEDICAL SCIENCES Year:2020 | Volume:8 | Issue:4 (29) Page(s): 73-87

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Title

Bioinformatic Design and Construction of a Chimeric Gene Comprising the stxB, ctxB and blf1 Genes

Author(s)

MOSHIRI ZAHRA | HADDAD RAHEEM | HONARI HOSSEIN | Issue Writer Certificate

Keywords

Burkholderia pseudomalleiQ1

BLF1 toxinQ1

MelioidosisQ1

Abstract

Introduction: bacteria species; Burkholderia pseudomallei, Vibrio cholera, and Shigella dysenteriae are the main pathogen causing Melioidosis, Cholera, and Shigella in humans respectively. bacterial species have a protein that has antigenic prosperities along with being a mucus based adjuvant thus easing their delivery and immunogenicity. These proteins were BLF1(B. pseudomallei), CTxB (V. cholera), and StxB (S. dysenteriae). The goal of this study was a chimeric gene composed of Blf1, CTxB, and StxB and expression of these proteins from E. coli. Materials and Methods: Using Bioinfoamtic tools such as Protparam and Modeller the different combinations of the chimeric protein were assessed. The blf1 and stxB genes were PCR amplified from pET28a-BFL1-StxB. The 850bp fragment encompassing BFL1-StxB was cloned using SalI and NotI in the pET28a-CTxB vector. The engineered plasmid was transformed in BL21(DE3). The ctxB-stxB-blf1 gene was expressed following induction with IPTG. This induction was then assessed using SDS-PAGE. Results: Chimeric protein instability index gets 40. 06 and the half-life of it in E. coli was over 10 hours. Codon Adaptive Index (CAI) rise to 0/9 and GC content increase to 54. 9. The optimal vaccine combination for the chimeric protein was determined to be CTxB-BLF1-StxB. Expression of recombinant protein in E. coli led to the production of a chimeric protein with 48 kDa molecular weight. Conclusion: The findings of the current study revealed that this antigen can be raised as a recombinant vaccine candidate which can be due to such factors as a property of the ability to stop protein synthesis in BLF1, CTxB adjuvant, and delivery of StxB protein.

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APA: Copy

MOSHIRI, ZAHRA, HADDAD, RAHEEM, & HONARI, HOSSEIN. (2020). Bioinformatic Design and Construction of a Chimeric Gene Comprising the stxB, ctxB and blf1 Genes. JOURNAL OF NEYSHABUR UNIVERSITY OF MEDICAL SCIENCES, 8(4 (29) ), 73-87. SID. https://sid.ir/paper/415255/en

Vancouver: Copy

MOSHIRI ZAHRA, HADDAD RAHEEM, HONARI HOSSEIN. Bioinformatic Design and Construction of a Chimeric Gene Comprising the stxB, ctxB and blf1 Genes. JOURNAL OF NEYSHABUR UNIVERSITY OF MEDICAL SCIENCES[Internet]. 2020;8(4 (29) ):73-87. Available from: https://sid.ir/paper/415255/en

IEEE: Copy

ZAHRA MOSHIRI, RAHEEM HADDAD, and HOSSEIN HONARI, “Bioinformatic Design and Construction of a Chimeric Gene Comprising the stxB, ctxB and blf1 Genes,” JOURNAL OF NEYSHABUR UNIVERSITY OF MEDICAL SCIENCES, vol. 8, no. 4 (29) , pp. 73–87, 2020, [Online]. Available: https://sid.ir/paper/415255/en

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