Molecular Characterization of Gene Encoding Outer Membrane Protein in Pathogenic Serovars in Iran.

The protein is highly conserved among pathogenic serovars and it is expressed during both acute and chronic infections. The aim of this study was to clone and sequence of the protein-encoding gene of serovars. In this study, 23 pathogenic serovars and two nonpathogenic serovars were used. These serovars were obtained from the microbial culture collection of Reference Laboratory, Department of Microbiology, Razi Vaccine and Serum Research Institute, Karaj, Iran. Three serovars, including Sejroe Hardjo-bovis, Grippotyphosa, Canicola, are used in the preparation of the trivalent vaccine. The gene was amplified by specific primers and the PCR products were then purified using kit and were cloned into a pTZ57R/T vector and transformed in competent DH5 cells. The cells were then plated onto LB agar containing ampicillin and recombinant colonies subjected to colony PCR to confirm the presence of the l gene. Positive colonies plasmid vector was isolated from cells by High Pure Plasmid Isolation Kit. The gene was detected in all 23 pathogenic serovars, while this gene was not observed in nonpathogenic It was determined that the similarity percentage of the sequenced pathogenic serovars is between 95.5% and 100%. The results concluded that the gene was highly conserved among various pathogenic serovars and can be used to develop an effective recombinant vaccine.