Cloning and expression of 51-kDa antigenic protein of Neorickettsia risticii NR-JA1
Park, M.K; Kim, E.H.; Cho, M.R.; Yi, Y.H.; Lee, M.J.; Shah, D.H.; Park, J.H.; Park, B.K.; Eo, S.K.; Lee, J.H. and Chae, J.S. (2005) Cloning and expression of 51-kDa antigenic protein of Neorickettsia risticii NR-JA1. Annals of the New York Academy of Sciences, 1063. pp. 246-251.
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Neorickettsia (Ehrlichia) risticii is a causative agent of acute diarrheal syndrome in horses, commonly known as Potomac horse fever. Korean isolate of N. risticii NR-JA1 was cultivated in mouse macrophage cell line P388D1. A complete ORF of p51 antigenic protein gene was amplified and cloned into pQE32 and pcDNA3.1 vectors and the resultant clones were named as pQE32/Nr-51 and pcDNA3.1/Nr-51, respectively. Recombinant p51 (rp51) protein antigen was expressed in E. coli (pQE32/Nr-51) and cos-7 cell line (pcDNA3.1/Nr-51). The rp51 protein showed immunoreactivity with anti- mouse p51 antibodies. BALB/c mice were inoculated with recombinant plasmid DNA (pcDNA3.1/Nr-51). The serum samples collected from these BALB/c mice showed IgG ELISA titers of 1:128. In a Western immunoblot assay, these serum samples showed a strong reactivity to rp51 expressed in cos-7 cell line transfected with pcDNA3.1/Nr-51. The results of this preliminary indicate that N. risticii p51 protein is an immmuno-dominant antigen and may be a good target for the development of serological or a molecular diagnostic test and possibly an improved recombinant DNA based vaccine against Potomac horse fever.
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