Use of a recombinant protein containing major epitopes of hnRNP G to detect anti-hnRNP G antibodies in dogs with systemic lupus erythematosus.
Res Vet Sci
; 81(3): 335-9, 2006 Dec.
Article
em En
| MEDLINE
| ID: mdl-16677675
ABSTRACT
The objective of this study was to express major epitopes of heterogeneous nuclear ribonucleoprotein G (hnRNP G) for detecting anti-hnRNP G antibodies in dogs with systemic lupus erythematosus (SLE). HnRNP G cDNA clone was isolated from HEp-2 cells, and a DNA fragment encoding immunodominant region (residues 189-272) of hnRNP G (hnRNP Gi) was subcloned into pET32 vector to construct a prokaryotic expression plasmid named pEThnRNPGi. After induction, Escherichia coli carrying pEThnRNPGi expressed a recombinant protein of 28 kDa, comprising recombinant hnRNP Gi and fusion tag. Purified recombinant hnRNP Gi protein was further analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and its identity was confirmed. Western blot analysis showed that recombinant hnRNP Gi was specifically recognized by anti-hnRNP G positive sera of SLE dogs, and not by negative control sera. In conclusion, recombinant hnRNP Gi protein expressed in this study may serve as a useful reagent to assist in the immunological diagnosis of canine SLE.
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Bases de dados:
MEDLINE
Assunto principal:
Proteínas Recombinantes
/
Epitopos Imunodominantes
/
Ribonucleoproteínas Nucleares Heterogêneas
/
Doenças do Cão
/
Lúpus Eritematoso Sistêmico
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
Idioma:
En
Revista:
Res Vet Sci
Ano de publicação:
2006
Tipo de documento:
Article