[Homology of the recombinant DNA of plasmid pDJH2 with the recombinant DNA probe of L. alstoni and analysis of its expression in Escherichia coli].
Dig-labeled recombinant DNA probe of L. alstoni which contains the entire structural OmpL1 gene was hybridized with the recombinant DNA of the plasmid named pDJH2 of the gene library of L. interrogans serovar lai strain 017. The result showed a high degree of homology among them; expression of recombinant DNA of pDJH2 was achieved by beta-D-galactosidase (IPTG) induction in E. coli. The molecular weight of this product is 68kd. Then they were treated with proteinase K and subjected to SDS-PAGE. The results showed it is a protein in nature. Using the specific monoclonal antibody E4B7D5 on immunoblotting and specific polyclonal antibody on dot-ELISA assay, we investigated the immune reaction and noticed that protein 68kd might be an antigen in character. E. coli which contains the recombinant plasmid pDJH2 were injected into BALB/c mice. Then the mice were challenged by leptospires of the strong virulence strain 017, but all the infected mice survived. In this paper, we first report the expression of recombinant DNA of L. interrogans serovar lai strain 017 in E. coli when injected with IPTG, and immunoprotection of BALB/c mice which were injected with the expression against the infection of L. interrogans serovar lai strain 017. pDJH2 may be the first recombinant for which the gene has been cloned and its expression product 68kd may be the immunoprotective antigens.
Jiang
N
N
Chengdu Institute of Biological Product, Ministry of Public Health.
Dai
B
B
Li
S
S
Zhao
H
H
Liu
J
J
Song
S
S
Yang
Y
Y
Zhang
Y
Y
Tu
Y
Y
Yang
H
H
D000818:Animals; D000942:Antigens, Bacterial / Q000096:biosynthesis* / Q000627:therapeutic use; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics*; D004274:DNA, Recombinant / Q000235:genetics*; D004926:Escherichia coli / Q000378:metabolism; D015723:Gene Library; D007919:Leptospira / Q000235:genetics*; D007921:Leptospira interrogans / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D012689:Sequence Homology, Nucleic Acid; D014895:Weil Disease / Q000517:prevention & control
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; D004274:DNA, Recombinant