Sequencing of the complete gene coding for the GroEL of the Wolbachia of Dirofilaria immitis and expression and purification of the recombinant protein.
Wolbachia are intracellular bacteria that infect arthropods and filarial nematodes. These bacteria play an important role in the immunology and pathogenesis of filarial diseases through their proteins and, possibly, other molecules. GroEL is a constitutively expressed bacterial protein; it is highly conserved among bacteria and is involved in the correct folding of newly synthesized proteins. Here we report the production of recombinant GroEL from the Wolbachia of Dirofilaria immitis. Our goal is to test the hypothesis that GroEL is involved in the immunopathology of filariases. The complete groel gene was PCR-amplified, sequenced and cloned into an expression vector. The recombinant GroEL was purified by affinity chromatography by using high-performance liquid chromatography (HPLC).
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Dipartimento di Patologia Animale, Igiene e Sanità Pubblica Veterinaria, Sezione di Patologia Generale e Parassitologia, Università degli Studi di Milano, Italy.
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Kramer
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D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000302:isolation & purification; D018834:Chaperonin 60 / Q000235:genetics* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D004269:DNA, Bacterial / Q000235:genetics; D004183:Dirofilaria immitis / Q000382:microbiology*; D005260:Female; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D020577:Wolbachia / Q000235:genetics* / Q000302:isolation & purification
D001426:Bacterial Proteins; D018834:Chaperonin 60; D004269:DNA, Bacterial; D011993:Recombinant Fusion Proteins