Cloning and sequencing of the gene present in , a surface protein in pathogenic leptospires.

BACKGROUND AND OBJECTIVES

Leptospirosis, an infection caused by pathogenic leptospires, is associated with insufficient sanitation and poverty. Leptospira is transmitted through contact with contaminated urine of reservoir animals. The primary objective of this study was to clone and sequence the gene present in local and vaccine serovars.

MATERIALS AND METHODS

A total of 16 serovars were cultured in EMJH liquid medium. After growing, genomic DNA was extracted using phenol-chloroform method. Primer pair was synthesized to amplify the 996 bp sequence. The amplified gene was cloned into pTZ57R/T vector. The sequences obtained from this study were compared with an only recorded sequence in the Genbank by the Meg Align software.

RESULTS

PCR products showed an amplified 996bp gene product belonging to pathogenic serovars, while no products were amplified in non-pathogenic serovars. Sequences comparison tests from 16 native serotypes examined in this study displayed a similarity range of 84% to 99.5% among serovars used. The results showed that two serotypes of including Serjoehardjo (RTCC2810 and RTCC2821) had the highest identity up to 95.5%. Two serovars of including Pomona (RTCC2822) and Icterohaemorrhagiae (RTCC2823) had the lowest identity about 84%.

CONCLUSION

As the results showed, , present on the surface of such bacteria, showed a conserved sequence. , as a key role in cell adhesion and pathogenicity, can be used for designing diagnostic tests and vaccines. Furthermore, sequencing of various sites in gene, including binding sites and immunogenic epitopes, can be valuable alternatives for future studies.