OPTIMIZATION OF GENE EXPRESSION AND PURIFICATION OF ENTEROTOXIGENIC ESCHERICHIA COLI RECOMBINANT LTB PROTEIN AND ANTIBODY PRODUCTION AGAINST IT

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KHALESI RAZIEH; NAZARIAN SH.; EHSAEI Z.; MANSOURI MEYSAM; AMANI J.; SALIMIAN J.; MOAZENI S.M.

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Journal: KOWSAR MEDICAL JOURNAL Year:2010 | Volume:15 | Issue:3 Page(s): 141-147

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Title

OPTIMIZATION OF GENE EXPRESSION AND PURIFICATION OF ENTEROTOXIGENIC ESCHERICHIA COLI RECOMBINANT LTB PROTEIN AND ANTIBODY PRODUCTION AGAINST IT

Author(s)

Keywords

ENTEROTOXIGENIC ESCHERICHIA COLIQ2

HEAT-LABILE TOXIN B SUBUNITQ3

EXPRESSIONQ3

RECOMBINANT VACCINEQ3

ANTIBODYQ2

Abstract

Aims: It has been estimated that gastroenteritis is caused by bacteria in 30-70% of cases. ENTEROTOXIGENIC ESCHERICHIA COLI or ETEC is one of the most common agents causing diarrhea. Protective immunity may be induced against this disease. Designing and producing a vaccine against this disease is one of the purposes of World Health Organization. Vaccine candidate molecules have to induce protective immunity against a broad spectrum of ETEC bacteria. Most ETEC strains can produce labile toxin; therefore labile toxin may be a proper candidate for being used as a vaccine molecule. The aim of this study was to optimize HEAT-LABILE TOXIN B SUBUNIT EXPRESSION in order to investigate its immunological properties.Materials & Methods: Optimizing of 3 parameters (IPTG concentration, time and temperature of promoter induction) was performed. Recombinant protein was purified with Ni-NTA column. Purified HEAT-LABILE TOXIN B SUBUNIT was injected to mice subcutaneously in 4 sessions. Blood samples were taken during the interval between the injections and after last injection. Then, ELISA was performed.Results: The optimum EXPRESSION occurred at 1mM IPTG concentration, after 3 hours and at 37oC. Recombinant protein was highly purified (>95%) with Ni-NTA column. Also, ELISA showed high titer of ANTIBODY production in mice.Conclusion: Expressed HEAT-LABILE TOXIN B SUBUNIT is an immunogenic protein and can be one of the important components in vaccine development against ETEC.

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APA: Copy

KHALESI, RAZIEH, NAZARIAN, SH., EHSAEI, Z., MANSOURI, MEYSAM, AMANI, J., SALIMIAN, J., & MOAZENI, S.M.. (2010). OPTIMIZATION OF GENE EXPRESSION AND PURIFICATION OF ENTEROTOXIGENIC ESCHERICHIA COLI RECOMBINANT LTB PROTEIN AND ANTIBODY PRODUCTION AGAINST IT. KOWSAR MEDICAL JOURNAL, 15(3), 141-147. SID. https://sid.ir/paper/32742/en

Vancouver: Copy

KHALESI RAZIEH, NAZARIAN SH., EHSAEI Z., MANSOURI MEYSAM, AMANI J., SALIMIAN J., MOAZENI S.M.. OPTIMIZATION OF GENE EXPRESSION AND PURIFICATION OF ENTEROTOXIGENIC ESCHERICHIA COLI RECOMBINANT LTB PROTEIN AND ANTIBODY PRODUCTION AGAINST IT. KOWSAR MEDICAL JOURNAL[Internet]. 2010;15(3):141-147. Available from: https://sid.ir/paper/32742/en

IEEE: Copy

RAZIEH KHALESI, SH. NAZARIAN, Z. EHSAEI, MEYSAM MANSOURI, J. AMANI, J. SALIMIAN, and S.M. MOAZENI, “OPTIMIZATION OF GENE EXPRESSION AND PURIFICATION OF ENTEROTOXIGENIC ESCHERICHIA COLI RECOMBINANT LTB PROTEIN AND ANTIBODY PRODUCTION AGAINST IT,” KOWSAR MEDICAL JOURNAL, vol. 15, no. 3, pp. 141–147, 2010, [Online]. Available: https://sid.ir/paper/32742/en

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