Vaccine Research
Vaccine Research
vacres
Medical Sciences
http://vacres.pasteur.ac.ir
1
admin
2383-2819
2423-4923
10.52547/vacres
en
jalali
1395
8
1
gregorian
2016
11
1
3
3
online
1
fulltext
fa
Cloning and expression of porA gene as the first step of a vaccine candidate study against Neisseria meningitidis serogroup A infection
Original article
Original article
<p dir="ltr" style="text-align: justify;"><strong>Introduction:</strong> <em>Neisseria meningitidis</em> is a major causative agent of bacterial septicemia and meningitis in human. PorA is a major component of the outer membrane of <em>N. meningitidis</em> and functions as a cationic Porin. This study aimed to clone and determine the expression of <em>PorA</em> as the first step for producing a proper antigen in a vaccine study against N. meningitidis. <strong>Methods:</strong> An approximately 1200-bp fragment of <em>porA</em> gene was amplified by PCR using <em>N. meningitidis</em> serogroup A genomic DNA and then cloned into prokaryotic expression vector pET-28a. The resulting construct (pET28a-<em>porA</em> plasmid) was transformed into competent <em>E.coli</em> BL21 cells for expression of recombinant protein. The proper overexpression of the recombinant protein was verified by SDS-PAGE and Western Blotting. <strong>R</strong><strong>esults: </strong>Cloning of <em>porA</em> was confirmed by colony-PCR and enzymatic digestion. The nucleotide sequence homology of the cloned <em>porA</em> gene was 97% , compared to the reference gene (NCBI GenBank accession number AL157959.1). The prokaryotic expression system (pET28a-<em>porA</em>- BL21) could produce our 45-kDa target recombinant protein, efficiently. <strong>Conclusion: </strong>The prokaryotic expression system and conditions used in this study provides an applicable method for producing recombinant <em>PorA</em> and possibly many other bacterial outer membrane proteins for future vaccine studies.</p>
Neisseria meningitidis, PorA, pET-28a.
60
63
http://vacres.pasteur.ac.ir/browse.php?a_code=A-10-1-30&slc_lang=fa&sid=1
P
Afrough
afroughparviz9@gmail.com
10031947532846002133
10031947532846002133
No
Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran. Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran.
Pasteur Institute of Iran. Tehran, Iran.
A
Behrouzi
10031947532846002134
10031947532846002134
No
Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran. Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran.
M
Davari
10031947532846002135
10031947532846002135
No
Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran.
MA
Malekan
10031947532846002136
10031947532846002136
No
Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran.
A
Fateh
afateh2@gmail.com
10031947532846002137
10031947532846002137
No
Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran. Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran.
F
Vaziri
farzam_vaziri@yahoo.com
10031947532846002138
10031947532846002138
No
Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran. Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran.
SD
Siadat
d.siadat@gmail.com
10031947532846002139
10031947532846002139
Yes
Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran. Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran.