The 6th International Congress of Clinical Laboratory and Clinic Cloning and Expression of FHBP Protein of Neisseria meningitidis in E. coli Yarian, F.*, Bandehpour, M.1, Kazemi, B.1 ,Ahangarzadeh, Sh.1 Shahid Beheshti University of Medical Sciences, Medical college, Biotechnology Department * f.yarian@gmail.com Background and Objective: Meningitis is the inflammation or infection of the membranes that is covering the brain or spinal cord. Bacterial meningitis have seen in children older than five years and it is typically more dangerous than virus and it can cause secondary problems in many cases in brain and nervous system. If it is diagnosed and treated early, Approximately ninety percent of children recover. Most common cause of bacterial meningitis in children is" Neisseria meningitidis " . Now if you suspect meningitis, even before laboratory results become available starting treatment with antibiotics. Fig1-Vector containing a FHBP gene construct Fig2-PQE 30 Vectore Material and methods: M Cut Un cut FHBP In this study, A DNA encoding the FHBP of Neisseria meningitidis was amplified by PCR using specific primers with appropriate restriction enzyme sites in their ends. The gene was cloned in the pQE30 vector and transformed to E. coli strain M15 and expressed in optimal condition. 3000 bp 860 bp 1000 bp 860 bp Findings: 470 bp 500 bp The desired gene is 860 nucleotides and it was Successfully PCR and cloned in pQE30 vector with enzymatic digestion, and it was confirmed by sequencing with universal primers and sequences were obtained for final approval. Protein obtained were approved by SDS-PAGE and Western blotting. This protein can be valuable for diagnosis of Meningitis Fig3-PCR:FHBP gene Fig4-digestion of vectore with Sal I & kpn I Fig5-Cloning confirmed by restriction enzyme digestion BL21 t0 t3 t5 t3 t5 M BL21 Conclusion: 70 kd We well cloned and expressed FHBP protein of Neisseria meningitidis in E. coli. The expressed protein was analyzed on SDS-PAGE and confirmed by western blotting using specific antibody to His tag. This protein can be valuable for diagnosis of Meningitis. 40 kd 32 kd Key words: Neisseria meningitidis bacteria, FHBP protein, cloning Fig6-SDS-page:expression of protein in BL21 (0.5 mM IPTG) Fig7-Western blotting: Confirm the protein Expression With anti his tag antibody Acknowledgment: I want to express my gratitude to Shahid Beheshti University of Medical science and center of cellular and molecular researches of Shahid Beheshti’s Medical Sciences for suporting this research and also wish to extend my thanks to Dr. M. Bandehpour, Dr. B.Kazemi, Dr .Z.Sharifnia, Dr. Sh. Ahangarzadeh, Miss. A. Koochaki and Miss. Z.Soleimanifar for their helpful comments. References: 1- Epidemic meningitis, meningococcaemia, and Neisseria meningitides- David S Stephens, Brian Greenwood, Petter Brandtzaeg 2-TZ YL, Martin LE,.Environmental survival of Neisseria meningitidis,Epidemol infect. 2014 Jan;142(1):187-90 3- Feibig T, Berti F Functional expression of the capsule polymerase of Neisseria meningitidis serogroup X: A new perspective for vaccine development.Glicobiology. 2014 Feb;24(2):150-8 Poster code:PBN-014