Abstract

Pseudomonas aeruginosa is one of the most important pathogens that causes many complications and deaths in the world every year. Many species of Pseudomonas aeruginosa have pili that are involved in the pathogenesis of the organism. The aim of this study was to clone PilF and PilQ genes of Pseudomonas aeruginosa in Escherichia coli and express it by real time PCR. Materials and Methods: After production of PCR product by Taq DNA polymerase and its cloning in PTG19-T cloning vector, then the vector was transduced to XL1-Blue host. After transformation, evaluation of the extracted RNA and amplification was performed by real-time PCR. Results: The analysis showed that the transfection of PilF and PilQ genes in PTG-T19 plasmid and Escherichia coli XL1-blue strain leads to high mRNA expression of these genes, which indicates the success of the proposed transducing method. TA cloning is a rapid and efficient method for transfecting the aforementioned genes as compared to conventional cloning methods. Due to the important role of PilF and PilQ genes, they can be used as a good candidate for developing the recombinant vaccines against infections caused by Pseudomonas aeruginosa.

Keywords: Cloning, Pseudomonas aeruginosa, PilF, PilQ, Antibiotic resistance