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Information Journal Paper

Title

CLONING, EXPRESSION AND PURIFICATION OF OUTER MEMBRANE SECRETIN PILQ406-770 OF NEISSERIA MENINGITIDIS SEROGROUP B

Pages

  193-199

Abstract

 Background: NEISSERIA MENINGITIDIS is a major cause of bacterial septicemia and meningitis. Currently, there are no vaccines to prevent disease caused by strains of N. meningitidis serogroup B, since cross-reactivity of the serogroup B capsule with human tissue has hampered efforts to develop a reliable vaccine. PilQ is an antigenically conserved outer membrane protein which is essential for meningococcal pilus EXPRESSION at the cell surface.Materials and methods: In the current study, we selected a 1095bp fragment of C-terminal of secretin pilQ and evaluated the immunogenicity of this recombinant fragment. This fragment was amplified by PCR from genomic DNA isolated from N. meningitidis serogroup B and cloned into the pET-28a EXPRESSION vector. PilQ406-770 was overexpressed with IPTG and then affinity-purified by Ni2+-Sepharose resin. The recombinant PilQ406-770 was reacted with rabbit anti- N.meningitidis polyclonal antibody in western-blot analysis. Mice were immunized subcutaneously with purified rPilQ 406-770 mixed with an equal volume of Freund's adjuvant and evaluated specific serum antibody responses.Results: Our results show pilQ 406-770 cloned in pET28a vector, while the CLONING of pilQ 406-770 was confirmed by colony- PCR and enzymatic digestion. SDS-PAGE analysis showed that our constructed prokaryotic EXPRESSION system pET28apilQ406- 770-BL21efficiently produces target RECOMBINANT PROTEIN with molecular weight of 43 kDa in the form of dissoluble inclusion body.Conclusion: Our results confirmed that a prokaryotic EXPRESSION system for PilQ406-770 protein was successfully constructed.

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    APA: Copy

    HAGHI, FAKHRI, NAJAR PERAYEH, SHAHIN, SIADAT, SEYED DAVAR, & MAHDAVI, MEHDI. (2010). CLONING, EXPRESSION AND PURIFICATION OF OUTER MEMBRANE SECRETIN PILQ406-770 OF NEISSERIA MENINGITIDIS SEROGROUP B. IRANIAN JOURNAL OF CLINICAL INFECTIOUS DISEASES, 5(4), 193-199. SID. https://sid.ir/paper/313664/en

    Vancouver: Copy

    HAGHI FAKHRI, NAJAR PERAYEH SHAHIN, SIADAT SEYED DAVAR, MAHDAVI MEHDI. CLONING, EXPRESSION AND PURIFICATION OF OUTER MEMBRANE SECRETIN PILQ406-770 OF NEISSERIA MENINGITIDIS SEROGROUP B. IRANIAN JOURNAL OF CLINICAL INFECTIOUS DISEASES[Internet]. 2010;5(4):193-199. Available from: https://sid.ir/paper/313664/en

    IEEE: Copy

    FAKHRI HAGHI, SHAHIN NAJAR PERAYEH, SEYED DAVAR SIADAT, and MEHDI MAHDAVI, “CLONING, EXPRESSION AND PURIFICATION OF OUTER MEMBRANE SECRETIN PILQ406-770 OF NEISSERIA MENINGITIDIS SEROGROUP B,” IRANIAN JOURNAL OF CLINICAL INFECTIOUS DISEASES, vol. 5, no. 4, pp. 193–199, 2010, [Online]. Available: https://sid.ir/paper/313664/en

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