title: Use of a recombinant protein containing major epitopes of hnRNP G to detect anti-hnRNP G antibodies in dogs with systemic lupus erythematosus.
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.
authors: Graduate Institute of Veterinary Microbiology, National Chung Hsing University, 250 Kuo Kuang Road, Taichung 40227, Taiwan, ROC.
authors: T-Y
authors: TY
authors: Lin
authors: L-C
authors: LC
authors: Chan
authors: Y-H
authors: YH
authors: Fan
authors: C-H
authors: CH
authors: Lin
authors: K-C
authors: KC
authors: Chow
authors: S-L
authors: SL
authors: Lin
authors: J-L
authors: JL
authors: Lan
authors: F-J
authors: FJ
authors: Lin
authors: S-H
authors: SH
authors: Chiou