Volume 24, Issue 102, March - April, 2020

Cloning of PilF & PilQ genes of Pseudomonas aeruginosa in Escherichia coli and expression by real time PCR

Zohreh Nasrabadi¹, Mitra Salehi^2♦, Kumars Amini³, Ahmad Majd⁴

¹Department of Biology, Faculty of Science, Islamic Azad University, North Tehran branch, Tehran, Iran
²Department of Microbiology, Faculty of Science, Islamic Azad University, North Tehran branch, Tehran, Iran
³Department of Microbiology, Faculty of Agriculture, Islamic Azad University, Saveh branch, Saveh, Iran
⁴Department of Biology, Faculty of Science, Islamic Azad University, North Tehran branch, Tehran, Iran

^♦Corresponding author
Department of Microbiology, Faculty of Science, Islamic Azad University, North Tehran branch, Tehran, Iran Email: drmitrasalehi@yahoo.com

ABSTRACT

Pseudomonas aeruginosa has emerged as one of the most common causes of hospital-acquired infections among patients with burn wounds, cystic fibrosis, acute leukemia, organ transplants, and intravenous-drug addiction, resulting in considerable annual mortality rates. Because of biofilm formation and its ability of rapidly acquires of resistance to many antibiotics, P. aeruginosa related infections are difficult to treat, and therefore, developing an effective vaccine is the most promising method for combating infection. In the present study, we aimed to transfect PilF and PilQ genes, essential genes to participate in the assembly and regulation of the type-4 pilus system, into E. coli strain XL-1 blue. The results of the present study revealed that transfection of PilF and PilQ genes in the PTG-T19 plasmid and E. coli XL1-blue strain resulted in the elevation of the mRNA expression level of these genes, suggestive of the success of our transducing method. Moreover, we suggest that TA cloning is a rapid and efficient method for transfecting the aforementioned genes as compared to conventional cloning methods. We also suggested for the first time that due to the presence of protected areas, PilF and PilQ could be considered as a good candidate for developing the recombinant vaccines against infections caused by Pseudomonas aeruginosa.

Keywords: Pseudomonas aeruginosa, PilF, PilQ, Recombinant vaccine, Antibiotic resistance.

Medical Science, 2020, 24(102), 606-612

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