Abstract

Background and aim: Acinetobacter baumannii is one of the emerging and important pathogens in various infections such as urinary tract infection, hospitals, meningitis and respiratory tract. The aim of this research is cloning and sequencing of ompA and smpA virulence genes of A. baumannii.

Methods: In this experimental study, the pathogenic A. baumannii was cultured on blood agar and macconkey agar mediums. Recognition of A. baumannii with microscopic, microbiologic and biochemical tests were performed. ompA and smpA genes were amplified by PCR and then cloned into the pTZ57R/T vector. The accuracy of the recombinant construct was carried out with PCR and enzymatic double digestion and then the both of ompA and smpA genes were sequenced. The evaluation of bacterial expression of these genes was performed with SDS-PSGE method in E. coli.

Result: The pathogenic A. baumannii was confirmed in biochemical and microbiological methods. After electrophoresis of the PCR products, the 1150 and 411 bp fragments of ompA and smpA genes were obtained, respectively. The results of PCR and double digestions on recombinant plasmids showed that the pTZ57R/T-ompA and pTZ57R/T-smpA were generated. The sequencing results confirmed the accuracy of the gene cloning.

Conclusion: The results showed that the pTZ57R/T is a suitable vector for the cloning of large fragment of PCR products and ompA and smpA are two highly conserved genes that appropriate for use as DNA vaccine.