Cloning, Expression and Purification of Outer Membrane Secretin PilQ406-770 of Neisseria Meningitidis Serogroup B
| Author | Fakhri Haghi | en |
| Author | Shahin Najar Perayeh | en |
| Author | Seyed Davar Siadat | en |
| Author | Mehdi Mahdavi | en |
| Issued Date | 2010-10-31 | en |
| 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 rPilQ406-770 mixed with an equal volume of Freund's adjuvant and evaluated specific serum antibody responses. Results: Our results show pilQ406-770 cloned in pET28a vector, while the cloning of pilQ406-770 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. | en |
| DOI | https://doi.org/ | en |
| Keyword | Cloning | en |
| Keyword | Expression | en |
| Keyword | Neisseria meningitidis | en |
| Keyword | PilQ406-770 | en |
| Keyword | Recombinant protein | en |
| Publisher | Brieflands | en |
| Title | Cloning, Expression and Purification of Outer Membrane Secretin PilQ406-770 of Neisseria Meningitidis Serogroup B | en |
| Type | Research Article | en |
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