The construction and expression of recombinant shuttle plasmid with ompl1 gene from leptosipra interrogans sereovar Lai 017 in bacilie calmette-guerin

摘要

Leptospirosis is a worldwide zoonosis caused by the pathogenic leptospires. In this study, we amplified the entire open reading frame of the OmpL1 gene from the genome of the leptospira servoar Lai strain 017. Two recombinant plasmids pBQ1 and pBQ2 were constructed by orientated ligation based on the E.coli-BCG shuttle plasmids pMV261 and pMV361 respectively. The recombinant plasmids were transformed into BCG by electroporation. The rBCGs bearing pBQ1and pBQ2 were induced by high temperature at 45C. The expessied product, a 35 KD protein was detected by SDS-PAGE. The result indicates that pBQ1 and pBQ2 can expess OmpL1 in rBCG. The technical methods in this study may help detect the immunogenicity and immunoprotection of OmpL1 and develop mor safe, highly effective rBCG bearing leptospiral antigen with long-lasting protection.