AMPLIFICATION AND CLONING OF A OUTER MEMBRANE PROTEIN (OMPL37) OF LEPTOSPIRA INTERROGANS SEROVAR CANICOLA, SERJOE HARDJO, GRIPPOTYPHOSA

Background and Aim:Leptospirosis is the most widespread zoonotic disease in the world, mainly in countries with humid subtropical or tropical climates. Outer membrane protein OmpL37 is an immunogenic protein which is present only in pathogenic serovars expressed during infection and it is conserved among different Leptospira serovars. In order to evaluate genetic conservation of the ompL37 gene, we cloned and sequenced this gene from Leptospira interrogans serovar Canicola, Serjoe hardjo, Grippotyphosa.Methods:Following the DNA extraction from the serovar, the ompL37 gene was amplified and cloned into pTZ57R/T vector and transformed into the competent E. coli (DH5α). Recombinant clones were confirmed by colony PCR and DNA sequencing. The related sequences were then analyzed and compared with the sequences in the Genbank databaseResults:PCR amplification of the ompL37 gene resulted in a 996 bp PCR product. The PCR based on the ompL37 gene detected all the pathogenic reference serovars of the tested Leptospira spp. In our study nucleotide sequencing results showed that the ompL37 has high identity between 84% and 99.5%. Vaccinal serovar Canicola(RTCC2805) exhibited 99.2% sequence identity with field serovars Canicola(RTCC2824,RTCC2836). There were 98.5% identity between vaccinal and field serovars of Grippotyphosa and 99.5% identity between vaccinal and field serovars of Serjoe hardjoConclusion:According to the results of this study and other researches, ompL37 gene was highly conserved among various pathogenic Leptospira serovars and can be cloned, expressed and used to develop an effective recombinant vaccine against leptospirosis. OmpL37 recombinant protein can also be used in an ELISA kit for the serodiagnosis of leptospirosis

Keywords:

Cloning , Leptospira , Outer membrane proteins , ompL37 gene