Cloning, sequencing, expression, and antigenic characterization of rMSP4 from Anaplasma marginale isolated from Paraná State, Brazil
Genet. mol. res. (Online)
;
6(1): 15-22, 2007. ilus
Article
Dans Anglais
| LILACS
| ID: lil-440617
ABSTRACT
Anaplasmosis is a bovine intraerythrocytic disease caused by the bacterium Anaplasma marginale; it causes significant economic losses in tropical and subtropical regions, worldwide. The msp4 gene of an A. marginale strain isolated in Paraná, Brazil, was amplified by PCR and sequenced; its cloning into the pET102/D-TOPO® vector produced an msp4-6xHis-V5-HP thioredoxin fusion gene construct. This recombinantclone was over-expressed in Escherichia coli BL21(DE-3); the expressed fusion protein was found almost entirely in the insoluble form (inclusion bodies) in the cell lysate. The inclusion bodies were solubilized with urea and the recombinant protein was purified by Ni-NTA column and dialyzed. This method produced a relatively high yield of rMSP4, which was used to immunize rabbits. The deduced amino acid sequence encoded by MSP4 showed 99% homology to A. marginale isolates from Florida, USA, and from Minas Gerais, Brazil. Both rMSP4 and native MSP4 were recognized by post- immunization rabbit serum, showing that rMSP4 has conserved epitopes. As antigenicity was preserved, rMSP4 might be useful for the development of vaccine against anaplasmosis.
Texte intégral:
Disponible
Indice:
LILAS (Amériques)
Sujet Principal:
Protéines bactériennes
/
Vaccins antibactériens
/
Anaplasma marginale
/
Protéines membranaires
/
Antigènes bactériens
Limites du sujet:
Animaux
Pays comme sujet:
Amérique du Sud
/
Brésil
langue:
Anglais
Texte intégral:
Genet. mol. res. (Online)
Thème du journal:
Biologie moléculaire
/
Génétique
Année:
2007
Type:
Article
Pays d'affiliation:
Brésil
Institution/Pays d'affiliation:
Universidade Estadual de Londrina/BR
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