Cloning and sequence analysis of the flagellin gene from Leptospira interrogans serovar Lai and serovar Hebdomadis
H.P.; Dal Baomin; Qiao Zhongdong; You Zili; Fang Zhimao
Journal of West China University of Medical Sciences 33(4): 518-521
Objective: To screen the candidate strain of Leptospiral nucleic acid vaccine and explain serovar differentiaiton. Methods: We amplified the endoflagellin flab gene from Leptospira interrograns serovar Lai strain 017 and serovar hebdomadis strain 56610 with PCR and cloned the flaB gene into the PGEM-T Easy Vector with the method of T/A directly cloning. Two recombinant plasmids (pDHTF017 and pDHTF610) were established by identification using PCR and restriction enzymes digestion. The two cloned flaB genes were sequenced with Sanger's DNA sequencing method. Results: The DNA sequencing result indicated that the flaB genes both contain 852bp and encode 283 amino acids. By analysis of the two flaB genes with softwere, it was found that their restriction sites are similar and inclucde many general restriction enzymes. Conclusion: Homologous analysis of DNA sequenc of the flaB gene from different serovars reveals that the homology between different serovars is very high (90%-99%).