{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2024,8,2]],"date-time":"2024-08-02T06:31:31Z","timestamp":1722580291831},"reference-count":0,"publisher":"Journal of Infection in Developing Countries","issue":"12","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J Infect Dev Ctries"],"abstract":"<jats:p>Introduction: Neisseria meningitidis is a major causative agent of bacterial septicemia and meningitis in humans. Currently, there are no vaccines to prevent disease caused by strains of N. meningitidis serogroup B. 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. Methodology: A 1200 bp fragment of porA gene was amplified by PCR from serogroup B N. meningitidis and then cloned into prokaryotic expression vector pET-32a. For expression of recombinant protein, pET32a-porA plasmid was transformed into competent Origami B (DE3) cells. Recombinant protein was overexpressed with isopropythio-beta-D-galctoside (IPTG) and affinity purified by Ni-NTA agarose. SDS-PAGE and western blotting were performed for protein determination and verification. Results: Cloning of porA was confirmed by colony-PCR and enzymatic digestion. In comparison with the corresponding sequences of original genes, the nucleotide sequence homology of the cloned porA gene was 97%. IPTG with a dosage of 1.0 mmol\/L could efficiently induce protein expression. SDS-PAGE analysis showed that our constructed prokaryotic expression system pET32a-PorA-Origami efficiently produces a target recombinant protein with a molecular weight of 65 kDa. The recombinant PorA was overexpressed as inclusion bodies and reacted with the serum from a rabbit previously immunized with native outer membrane vesicle. Conclusion: This prokaryotic expression system provides an easy method for producing recombinant PorA and may also be useful for the production of other bacterial outer membrane proteins for vaccine studies.<\/jats:p>","DOI":"10.3855\/jidc.1557","type":"journal-article","created":{"date-parts":[[2011,12,15]],"date-time":"2011-12-15T02:48:30Z","timestamp":1323917310000},"page":"856-862","source":"Crossref","is-referenced-by-count":9,"title":["Cloning, expression and purification of outer membrane protein PorA of Neisseria meningitidis serogroup B"],"prefix":"10.3855","volume":"5","author":[{"given":"Fakhri","family":"Haghi","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shahin","family":"Najar Peerayeh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seyed Davar","family":"Siadat","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mehran","family":"Montajabiniat","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"2222","published-online":{"date-parts":[[2011,11,14]]},"container-title":["The Journal of Infection in Developing Countries"],"original-title":[],"link":[{"URL":"https:\/\/jidc.org\/index.php\/journal\/article\/download\/22169784\/640","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/jidc.org\/index.php\/journal\/article\/download\/22169784\/640","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2019,7,16]],"date-time":"2019-07-16T00:26:55Z","timestamp":1563236815000},"score":1,"resource":{"primary":{"URL":"https:\/\/jidc.org\/index.php\/journal\/article\/view\/22169784"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2011,11,14]]},"references-count":0,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2011,12,13]]}},"URL":"https:\/\/doi.org\/10.3855\/jidc.1557","relation":{},"ISSN":["1972-2680"],"issn-type":[{"value":"1972-2680","type":"electronic"}],"subject":[],"published":{"date-parts":[[2011,11,14]]}}}