Identification and cloning of the oligopeptide binding protein (OPPA) Xanthomonas axonopodis pv citri (2003)
- Authors:
- USP affiliated authors: FERREIRA, RITA DE CASSIA CAFE - ICB ; FERREIRA, LUIS CARLOS DE SOUZA - ICB
- Unidade: ICB
- Assunto: MICROBIOLOGIA
- Language: Português
- Abstract: Uptake of oligopeptides with more than three amino acids is mediated by the oligopeptide uptake system, an ABC-type transport system composed of four membrane-bound docking elements (OppB, OppC, OppD, OppF) and one binding periplasmic component, the oligopeptide binding protein (OppA), which confer substrate affinity and specificity to the transport system. In this work the oppA orthologue of the plant pathogen Xanthomonas axonopodis pv. citri were cloned and expressed in Escherichia coli K12 BL21 cells using the pET28 and pET29 expression vectors. Recombinant proteins fused to poly-histidine tag sequences at both N- and C-terminal regions of the OppA protein were expressed at high levels by E. coli K12 strains transformed with the engineered plasmid vectors.All proteins were expressed as inclusion bodies in the cytoplasm of the recombinant proteins, which were purified either by affinity chromatography with Ni-containing resin or ion-exchange chromatography following solubilization of the insoluble proteins. Specific anti-OppA antibodies were raised in mice immunized with the purified recombinant OppA proteins and used to monitor expression of the OppA in periplasm-enriched fractions of X. auxonopodis pv citri during in vitro growth. Further analysis of the Xanthomonas OppA may help to define substrate affinity and specificity of this protein
- Imprenta:
- Publisher: Comissão de Pesquisa do ICB/USP
- Publisher place: São Paulo
- Date published: 2003
- Source:
- Título do periódico: Resumos
- Conference titles: Congresso Instituto Ciências Biomédicas, IV
-
ABNT
PACCEZ, Juliano Domiraci et al. Identification and cloning of the oligopeptide binding protein (OPPA) Xanthomonas axonopodis pv citri. 2003, Anais.. São Paulo: Comissão de Pesquisa do ICB/USP, 2003. . Acesso em: 19 abr. 2024. -
APA
Paccez, J. D., Ferreira, R. de C. C., Ferreira, L. C. de S., Suzuki, C. F., & Balan, A. (2003). Identification and cloning of the oligopeptide binding protein (OPPA) Xanthomonas axonopodis pv citri. In Resumos. São Paulo: Comissão de Pesquisa do ICB/USP. -
NLM
Paccez JD, Ferreira R de CC, Ferreira LC de S, Suzuki CF, Balan A. Identification and cloning of the oligopeptide binding protein (OPPA) Xanthomonas axonopodis pv citri. Resumos. 2003 ;[citado 2024 abr. 19 ] -
Vancouver
Paccez JD, Ferreira R de CC, Ferreira LC de S, Suzuki CF, Balan A. Identification and cloning of the oligopeptide binding protein (OPPA) Xanthomonas axonopodis pv citri. Resumos. 2003 ;[citado 2024 abr. 19 ] - Bacillus subtilis endospores at high purity and recovery yields: optimization of growth conditions and purification method
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