Cloning and sequencing of the virulent gene LipL32 of Leptospira interrogans serovar Autumnalis

Abstract

Aim: To clone the virulent gene LipL32 of Leptospira interrogans serovar Autumnalis and to analyze the sequence with LipL32 gene of other pathogenic serovars of Leptopsira. Materials and Methods: Leptospira interrogans serovar Autumnalis procured from Leptospira research laboratory, Chennai was used in the study. Polymerase chain reaction (PCR) was carried out for amplifying LipL32 gene using the reported primers of Leptospira Kirschnerii. The PCR product was cloned into TA cloning vector and the vector was transformed into E.Coli DH5á cells. The plasmid was isolated from E.Coli and sent for sequencing with universal primers. The sequence was submitted in genbank with accession number JQ861883. Results: The PCR product revealed an amplicon of 790 bp. The LipL32 gene sequence of Leptospira interrogans serovar Autumnalis showed 99 % similarity with most of the pathogenic Leptospires. Conclusions: LipL32 gene of Leptospira is highly conserved in most of the pathogenic Leptospires. The study concludes that this gene could be used as a target for the diagnosis of leptospirosis in animals and humans and could be tested as an important candidate antigen for vaccine production. [Vet World 2013; 6(4.000): 193-195]

Keywords

• cloning
• sequencing
• LipL32 gene
• Leptospira Autumnalis