CLONING AND EXPRESSION OF RECOMBINANT LIPL41 ANTIGEN FROM LEPTOSPIRA INTERROGANS IN PROKARYOTIC SYSTEM

Background and Aim:Leptospirosis is a worldwide zoonosis caused by pathogenic Leptospira spp. Outer membrane proteins of leptospire are among the most effective antigens which can stimulate remarkable immune responses during the infection processes. LipL41 is the third most abundant lipoprotein found in the outer membranes of pathogenic leptospires and has been considered a putative virulence factor. The objective of the present study was Cloning and expression of recombinant LipL41 antigen from Leptospira interrogans in prokaryotic system.Methods:LipL41 was clonned in E. coli strain BL21 using pET32 + plasmid as a vector and induced with 25 µM IPTG at 22 °C for 16 hours. The bacterial pellet was obtained by centrifugation (5,000 rpm). The resuspended cells were disrupted by sonication. The cell lysate was centrifuged at 15,000 rpm at 4 °C for 20 min. Samples were solubilized in sample buffer plus mercaptoenthaol , Proteins were separated on 10% sodium dodecyl sulfate (SDS) and followed by Coomassie Brilliant Blue staining.Results:The protein was successfully expressed in Escherichia coli BL21 and purified. SDS-PAGE results showed that the full-length 47kD protein was induced by IPTGConclusion:Leptospirosis is considered as a reemerging infectious disease, not only for the increase in its incidence during the past recent years but also for the increased severity of the illness. The cloned gene could be further used for expression of recombinant protein for serodiagnosis and leading candidate vaccine antigens of leptospirosis.

کلیدواژه ها:

Leptospira ، LipL41cloning ، expression