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:: دوره 3، شماره 7 - ( 11-1394 ) ::
جلد 3 شماره 7 صفحات 22-25 برگشت به فهرست نسخه ها
Cloning and expression of porA gene as the first step of a vaccine candidate study against Neisseria meningitidis serogroup A infection
چکیده:   (691 مشاهده)

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.

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نوع مطالعه: Original article |
دریافت: ۱۳۹۶/۱/۲۶ | پذیرش: ۱۳۹۶/۱/۲۶ | انتشار: ۱۳۹۶/۱/۲۶
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DOI: 10.18869/acadpub.vacres.3.7.22


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Afrough P, Behrouzi A, Davari M, Malekan M, Fateh A, Vaziri F et al . Cloning and expression of porA gene as the first step of a vaccine candidate study against Neisseria meningitidis serogroup A infection. vacres. 2016; 3 (7) :22-25
URL: http://vacres.pasteur.ac.ir/article-1-93-fa.html
Cloning and expression of porA gene as the first step of a vaccine candidate study against Neisseria meningitidis serogroup A infection. تحقیقات واکسن. 1394; 3 (7) :22-25

URL: http://vacres.pasteur.ac.ir/article-1-93-fa.html

دوره 3، شماره 7 - ( 11-1394 ) برگشت به فهرست نسخه ها
تحقیقات واکسن Vaccine Research
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