Parasites, Hosts and Diseases
- 제39권3호
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- Pages.241-246
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- 2001
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- 2982-5164(pISSN)
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- 1738-0006(eISSN)
Determination of antigenic domain in GST fused major surface protein (Nc-p43) of Neospora caninum
- Son, Eui-Sun (Department of Parasitology and Catholic Institute of Parasitic Diseases, Catholic University of Korea, College of Medicine) ;
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Ahn, Hye-Jin
(Department of Parasitology and Catholic Institute of Parasitic Diseases, Catholic University of Korea, College of Medicine) ;
- Kim, Jae-Hoon (National Veterinary Research and Quarantine Service) ;
- Kim, Dae-Yong (Department of Veterinary Pathology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University) ;
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Nam, Ho-Woo
(Department of Parasitology and Catholic Institute of Parasitic Diseases, Catholic University of Korea, College of Medicine)
- 발행 : 2001.09.01
초록
The antigenic domain of the major surface protein (Nc-p43) of Neospora caninum was examined by polymerase chain reaction of its gene fragments and recombinant expression as GST fusion proteins. The fragments of Nc-p43 were as follow: a total open reading frame (OFR), T: OFR without signal sequence and C-terminal hydrophobic sequence, S: N-terminal 2/3 parts of S, A: C-terminal 2/3 parts, P; N-terminal 1/3 part, X: middle 1/3 part Y; and C-terminal 1/3 part, Z, respectively. The DNA fragments were cloned into pGEX-47 vector. Recombinant plasmids transformed into Escherichia coli of BL21 pLysS (DE3) strain were induced to express GST or GST fused fragments of Nc-p43 such as 69 kDa protein for T,66 kDa for S, 52 kDa for A,53 kDa for P, and 40 kDa proteins for X, Y, and Z, respectively in SDS-PAGE. The Nc-p43 fragments of T, S, and P reacted with a bovine serum of neosporosis while those of A, X, Y, and Z together with GST did not in the western blot. These findings suggest that the antigenic domain of Nc-p43 of N. caninum may be localized in the C-terminal 2/3 parts. Together with Al9 clone in SAGI of Toxoplasma gondii (Nam et at., 1996), the P fragment of Nc-p43 could be used as efficient antigens to diagnose and differentiate those infections with both species .