Cloning and sequencing of LigB gene of Leptospira interrogans serovar Canicola

Primers were designed with built-in restriction sites to clone the immunogenic moiety of the LigB outer membrane gene of Leptospira interrogans serovar Canicola. The polymerase chain reaction (PCR) amplified product of LigB gene was cloned in pPROEXHTb vector and transformed into Escherichia coli DH5α cells. The positive recombinant clones were confirmed by colony Polymerase Chain Reaction (colony PCR), Restriction Endonuclease (RE) digestion and DNA sequencing. The sequenced gene was analysed for its homology with the already reported sequence of other Leptospira serovars and it revealed that serovar Canicola was closely related with serovar Grippotyphosa and distantly related with Pomona.