RECOMBINANT EXPRESSION OF STAPHYLOCOCCUS AUREUS ENTEROTOXIN TYPE B AS A VACCINE CANDIDATE

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HOSSEINI SEYED AMIR; EBRAHIMI FIROUZ; NAZARIAN SHAHRAM; HAMIDI MAHMUD

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Journal: Jundishapur Scientific Medical Journal Year:2017 | Volume:16 | Issue:6 Page(s): 653-664

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Title

RECOMBINANT EXPRESSION OF STAPHYLOCOCCUS AUREUS ENTEROTOXIN TYPE B AS A VACCINE CANDIDATE

Author(s)

HOSSEINI SEYED AMIR | EBRAHIMI FIROUZ | NAZARIAN SHAHRAM | HAMIDI MAHMUD | Issue Writer Certificate

Keywords

STAPHYLOCOCCUS AUREUSQ1

SEB TOXINQ1

RECOMBINANT EXPRESSIONQ1

WESTERN BLOTTINGQ2

VACCINE CANDIDATEQ1

Abstract

Background and Objective: STAPHYLOCOCCUS AUREUS is a gram-positive cocci that causes many diseases, such as food poisoning, risky shocks. Entrotoxins are the most important toxins of the bacteria. Among them, enterotoxin type B is the most common and important ones which is a super antigen. The aim of this study was the cloning and RECOMBINANT EXPRESSION of seb gene in order to achieve a stable construct for the protein production and further use as a VACCINE CANDIDATE in future investigations. Subjects and Methods: seb gene was analyzed for rare codons and gene optimaztion was performed using bioinformatic softwares. Enzymatic digestion was performed to confirm the presence of the seb gene into pET28a(+) expression vector. RECOMBINANT EXPRESSION of SEB was induced by IPTG following cloning confirmation. Then, protein purification was done under non-denaturing conditions using a nickel chromatography column. Protein confirmation was performed by WESTERN BLOTTING analysis. Results: Codon adaptation index (CAI) of the native gene was 0. 68, while the optimized sequence had a CAI of 0. 82. Percentage of codon having high frequency distribution was improved to 57%. Restriction analysis confirmed the cloning of the seb gene into pET28a vector. The results showed that the cloning of seb gene in pET28a(+) vector was performed in an appropriate position between expected restriction sites. In addition, SEB had a good and significant expression after induction by IPTG. The total yield of purified protein was 22mg/L. WESTERN BLOTTING showed specific reactivity of recombinant protein with anti-his tag antibody. Conclusion: Stable recombinant construct containing seb gene was constructed and appropriate RECOMBINANT EXPRESSION of the protein was observed. So, this protein could be used in future studies as a VACCINE CANDIDATE against SEB TOXIN of STAPHYLOCOCCUS AUREUS.

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APA: Copy

HOSSEINI, SEYED AMIR, EBRAHIMI, FIROUZ, NAZARIAN, SHAHRAM, & HAMIDI, MAHMUD. (2017). RECOMBINANT EXPRESSION OF STAPHYLOCOCCUS AUREUS ENTEROTOXIN TYPE B AS A VACCINE CANDIDATE. JUNDISHAPUR SCIENTIFIC MEDICAL JOURNAL, 16(6 ), 653-664. SID. https://sid.ir/paper/12482/en

Vancouver: Copy

HOSSEINI SEYED AMIR, EBRAHIMI FIROUZ, NAZARIAN SHAHRAM, HAMIDI MAHMUD. RECOMBINANT EXPRESSION OF STAPHYLOCOCCUS AUREUS ENTEROTOXIN TYPE B AS A VACCINE CANDIDATE. JUNDISHAPUR SCIENTIFIC MEDICAL JOURNAL[Internet]. 2017;16(6 ):653-664. Available from: https://sid.ir/paper/12482/en

IEEE: Copy

SEYED AMIR HOSSEINI, FIROUZ EBRAHIMI, SHAHRAM NAZARIAN, and MAHMUD HAMIDI, “RECOMBINANT EXPRESSION OF STAPHYLOCOCCUS AUREUS ENTEROTOXIN TYPE B AS A VACCINE CANDIDATE,” JUNDISHAPUR SCIENTIFIC MEDICAL JOURNAL, vol. 16, no. 6 , pp. 653–664, 2017, [Online]. Available: https://sid.ir/paper/12482/en

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