Expression, purification and characterization of surface proteins from Leptospira interrogans. / Expressão, purificação e caracterização de proteínas de superfície de Leptospira interrogans.
AUTOR(ES)
Marina von Atzingen dos Reis
DATA DE PUBLICAÇÃO
2009
RESUMO
The whole-genome sequences of L. interrogans serovar Copenhageni and the bioinformatic tools allow us to choose fifteen genes encoding for conserved hypothetical proteins predicted to be exported to the membrane. The chosen genes were amplified by PCR from six predominant pathogenic serovars, the DNA cloned in an E. coli vector, the recombinant proteins expressed in fusion with 6xHis-tag at N-terminus and purified by metal affinity chromatography. Six proteins were recognized by antibodies present in sera from human patients diagnosed with leptospirosis. By ELISA-attachment assay, we have identified a novel adhesion, named Lsa21, that binds strongly to laminin, collagen IV, and plasma fibronectin. By western blotting assay, we have further identified nine novel probable adhesions. The immunization/challenge assays showed that the recombinant protein rLIC12730 afforded protection against lethal leptospiral inoculation in hamsters. Our data suggest that it is a promising candidate for prevention of leptospirosis.
ASSUNTO(S)
biotecnologia bacteria genomas bactérias biotechnology genomes recombinant proteins vacinas leptospira leptospira proteínas recombinantes vaccines
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