CLONING, FUSION AND EXPRESSION OF CTB AND N-TERMINAL IPAD GENE CASSETTE IN E. COLI

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HONARI H.; GHOFRANI M.; SAADATI M.; MINAEI M.E.

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Information Journal Paper

Title

CLONING, FUSION AND EXPRESSION OF CTB AND N-TERMINAL IPAD GENE CASSETTE IN E. COLI

Author(s)

HONARI H. | GHOFRANI M. | SAADATI M. | MINAEI M.E. | Issue Writer Certificate

Keywords

ADJUVANTQ2

CLONINGQ2

CTB GENEQ2

FUSION GENEQ2

IPAD GENEQ2

SHIGELLOSISQ2

Abstract

Background: Having no effective vaccine, Shigella dysenteriae is of the main causes of diarrhea. IpaD protein is of the most important factors in high virulence of pathogenic shigella.Cloning of N-terminal IPAD GENEs along with CTB GENEs that possess ADJUVANT and carrier roles, as a recombinant vaccine, can enhance mucosal immune response.Materials and Methods: In an experimental study, primers of ctB and IPAD GENEs were designed based on gene cassette construction. PCR reactions were performed to propagate the fragments. Propagated fragments, then, were cloned into pGEM-Teasy vectors. Both vectors, then, were cut by restriction enzymes of HindIII and XhoI, and finally ipaD and ctB were fused together. In the next step, the ctB-ipaD gene cassette and expression vector of pET28a (+) were cut by SalI and HindIII restriction enzymes. Then, ctB-ipaD cassette was expresses in the expression vector.Results: In prest study, the N terminal of ipaD as a vaccine candidate antigen was genetically linked to the C terminal of ctB which possesses carrier and ADJUVANT potency. Fusion of ctB-ipaD in the expression vector pET28a (+) was confirmed by PCR and restriction enzymes digestion. The recombinant proteins produced then was confirmed by SDS-PAGE and Western blot.Conclusion: According to previous studies, production of ctB-ipaD cassette and its expression was the aim of this study. In hop for a vaccine against Shigella, protein expression of this gene cassette and production of an appropriate should be followed in future studies.

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APA: Copy

HONARI, H., GHOFRANI, M., SAADATI, M., & MINAEI, M.E.. (2014). CLONING, FUSION AND EXPRESSION OF CTB AND N-TERMINAL IPAD GENE CASSETTE IN E. COLI. ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ), 16(12 (81)), 0-0. SID. https://sid.ir/paper/68887/en

Vancouver: Copy

HONARI H., GHOFRANI M., SAADATI M., MINAEI M.E.. CLONING, FUSION AND EXPRESSION OF CTB AND N-TERMINAL IPAD GENE CASSETTE IN E. COLI. ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ)[Internet]. 2014;16(12 (81)):0-0. Available from: https://sid.ir/paper/68887/en

IEEE: Copy

H. HONARI, M. GHOFRANI, M. SAADATI, and M.E. MINAEI, “CLONING, FUSION AND EXPRESSION OF CTB AND N-TERMINAL IPAD GENE CASSETTE IN E. COLI,” ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ), vol. 16, no. 12 (81), pp. 0–0, 2014, [Online]. Available: https://sid.ir/paper/68887/en

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