MOLECULAR CHARACTERIZATION AND FUNCTIONAL ANALYSIS OF THE PILQ380-706: A NOVEL SECRETIN DOMAIN IN PSEUDOMONAS AERUGINOSA

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FAEZI SOBHAN; NIKOKAR IRAJ; ELMI ALI; GHASEMI YUSUF; FARAHBAKHSH MOJTABA; SALIMI CHIRANI ALIREZA; MAHDAVI MEHDI

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Journal: Avicenna Journal of Medical Biotechnology Year:2018 | Volume:10 | Issue:1 Page(s): 34-40

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Title

MOLECULAR CHARACTERIZATION AND FUNCTIONAL ANALYSIS OF THE PILQ380-706: A NOVEL SECRETIN DOMAIN IN PSEUDOMONAS AERUGINOSA

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Keywords

PSEUDOMONAS AERUGINOSA

PROTEINS

SECRETIN

Abstract

Background: Type 4 pili (T4P) is an important virulence factor of PSEUDOMONAS AERUGINOSA (P. aeruginosa). T4P pass the outer membrane through a large oligomeric channel made of a single PilQ protein that is most highly conserved at their C-termini. To develop a functional vaccine that can be used in clinical application, the SECRETIN domain of the PilQ (PilQ380-706) was produced as a recombinant protein.Methods: A 981 bp fragment of C-terminal of the pilQ SECRETIN (pilQ1138-2118) from was designed into the prokaryotic expression vector pET28a. The presence of the pilQ1138-2118 gene in the recombinant construct (pET28a/pilQ) was assessed by double digestion and PCR. After transformation, expression of the recombinant PilQ was induced by addition of IPTG. The expressed recombinant protein was purified by a modified method using a HisTrap affinity column and finally confirmed by SDS-PAGE. The functional activities of the produced PilQ380-706 confirmed by Western blot analysis and twitching inhibition assay.Results: The PCR and enzymatic digestion results showed the presence of the pilQ1138-2118 gene in the construct. The protein electrophoresis showed that the molecular weight of the recombinant PilQ380-706 is approximately 37 kDa. The Western blot analysis confirmed the specificity of specific IgG against the PilQ380-706 protein. The PilQ380-706 protein showed high biological activity in all of these standard assays.Conclusion: Since, the PilQ380-706 protein plays an important role in the biogenesis of pili; and thus, the primary establishment of P. aeruginosa; it seems that it can be used as a candidate vaccine or an adjuvant in the future studies.

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APA: Copy

FAEZI, SOBHAN, NIKOKAR, IRAJ, ELMI, ALI, GHASEMI, YUSUF, FARAHBAKHSH, MOJTABA, SALIMI CHIRANI, ALIREZA, & MAHDAVI, MEHDI. (2018). MOLECULAR CHARACTERIZATION AND FUNCTIONAL ANALYSIS OF THE PILQ_380-706: A NOVEL SECRETIN DOMAIN IN PSEUDOMONAS AERUGINOSA. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), 10(1), 34-40. SID. https://sid.ir/paper/313937/en

Vancouver: Copy

FAEZI SOBHAN, NIKOKAR IRAJ, ELMI ALI, GHASEMI YUSUF, FARAHBAKHSH MOJTABA, SALIMI CHIRANI ALIREZA, MAHDAVI MEHDI. MOLECULAR CHARACTERIZATION AND FUNCTIONAL ANALYSIS OF THE PILQ_380-706: A NOVEL SECRETIN DOMAIN IN PSEUDOMONAS AERUGINOSA. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB)[Internet]. 2018;10(1):34-40. Available from: https://sid.ir/paper/313937/en

IEEE: Copy

SOBHAN FAEZI, IRAJ NIKOKAR, ALI ELMI, YUSUF GHASEMI, MOJTABA FARAHBAKHSH, ALIREZA SALIMI CHIRANI, and MEHDI MAHDAVI, “MOLECULAR CHARACTERIZATION AND FUNCTIONAL ANALYSIS OF THE PILQ_380-706: A NOVEL SECRETIN DOMAIN IN PSEUDOMONAS AERUGINOSA,” AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), vol. 10, no. 1, pp. 34–40, 2018, [Online]. Available: https://sid.ir/paper/313937/en

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