%0 Journal Article %A Afrough, P %A Behrouzi, A %A Davari, M %A Malekan, MA %A Fateh, A %A Vaziri, F %A Siadat, SD %T Cloning and expression of porA gene as the first step of a vaccine candidate study against Neisseria meningitidis serogroup A infection %J Vaccine Research %V 3 %N 3 %U http://vacres.pasteur.ac.ir/article-1-93-en.html %R 10.18869/acadpub.vacres.3.7.22 %D 2016 %K Neisseria meningitidis, PorA, pET-28a., %X Introduction: Neisseria meningitidis is a major causative agent of bacterial septicemia and meningitis in human. PorA is a major component of the outer membrane of N. meningitidis and functions as a cationic Porin. This study aimed to clone and determine the expression of PorA as the first step for producing a proper antigen in a vaccine study against N. meningitidis. Methods: An approximately 1200-bp fragment of porA gene was amplified by PCR using N. meningitidis serogroup A genomic DNA and then cloned into prokaryotic expression vector pET-28a. The resulting construct (pET28a-porA plasmid) was transformed into competent E.coli BL21 cells for expression of recombinant protein. The proper overexpression of the recombinant protein was verified by SDS-PAGE and Western Blotting. Results: Cloning of porA was confirmed by colony-PCR and enzymatic digestion. The nucleotide sequence homology of the cloned porA gene was 97% , compared to the reference gene (NCBI GenBank accession number AL157959.1). The prokaryotic expression system (pET28a-porA- BL21) could produce our 45-kDa target recombinant protein, efficiently. Conclusion: The prokaryotic expression system and conditions used in this study provides an applicable method for producing recombinant PorA and possibly many other bacterial outer membrane proteins for future vaccine studies. %> http://vacres.pasteur.ac.ir/article-1-93-en.pdf %P 60-63 %& 60 %! %9 Original article %L A-10-1-30 %+ Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran. Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran. %G eng %@ 2383-2819 %[ 2016