The construction and expression of recombinant shuttle plasmid with OmpL1 gene from leptospira interrogans serovar Lai strain 017 in Bacille Calmette-Guerin.
OBJECTIVE
To construct recombinant BCG against leptospirosis.


METHODS
We amplified the entire open reading frame of the OmpL1 gene from the genome of the leptospire serovar Lai strain 017. Two recombinant plasmids pBQ1 and pBQ2 were constructed by oriented 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 pBQ1 and pBQ2 were induced by high temperature of 45 degrees C.


RESULTS
The expressed product, a 35kD protein was detected by SDS-PAGE. The result indicates that pBQ1 and pBQ2 can express OmpL1 in rBCG.


CONCLUSIONS
The technical methods in this study may help detect the immunogenicity and immunoprotection of OmpL1 and develop more safe, highly effective rBCG bearing leptospiral antigen with long-lasting protection.
Bao
Lang
L
Research Unit of Leptospirosis & Infection and Immunity, West China University of Medical Sciences, Chengdu 610041.
Qiu
Hongyu
H
Yan
Jufang
J
Xie
Yongen
Y
Chen
Wei
W
D001500:BCG Vaccine / Q000235:genetics*; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D007921:Leptospira interrogans / Q000235:genetics*; D016366:Open Reading Frames / Q000235:genetics*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis
D001500:BCG Vaccine; D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; D011994:Recombinant Proteins; C081904:ompL1 protein, Leptospira