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http://dx.doi.org/10.7853/kjvs.2014.37.3.143

Expression of porcine reproductive and respiratory syndrome virus (PRRSV) ORF7 gene and monoclonal antibody production  

Lee, Seung-Chul (Choongang Vaccine Laboratories)
Park, Ga-Hye (College of Veterinary Medicine, Chungbuk National University)
Lee, Kyeong-Won (College of Veterinary Medicine, Chungbuk National University)
Ryu, Min-Sang (College of Veterinary Medicine, Chungbuk National University)
Kang, Shien-Young (College of Veterinary Medicine, Chungbuk National University)
Publication Information
Korean Journal of Veterinary Service / v.37, no.3, 2014 , pp. 143-150 More about this Journal
Abstract
Porcine reproductive and respiratory syndrome virus (PRRSV) is the etiological agent of PRRS characterized by reproductive losses in sows and respiratory disorders in piglets. The PRRSV is a small enveloped virus containing a positive-sense, single-stranded RNA genome and divided into two genotype, type 1 (European) and type 2 (North American), respectively, by nucleotide identity. In this study, ORF7 gene of the type 1 and type 2 PRRSV was cloned and expressed in Baculovirus expression system. Also, monoclonal antibodies (MAbs) against ORF7 were produced and characterized. The expressed ORF7 proteins in the recombinant virus were confirmed by indirect fluorescence antibody (IFA) test using His6 and PRRSV-specific antiserum. A total of eight MAbs were produced and characterized. One (3G12) MAb was type 1 PRRSV ORF7-specific and two (6B10 and 16H8) were type 2 PRRSV ORF7-specific. Other five (1A1, 2A4, 4B4, 12C4 and 13F11) MAbs reacted with both type 1 and type 2 PRRSV. Some PRRSV ORF7-specific MAbs recognized the porcine tissues infected with PRRSV by IFA or immunohistochemistry (IHC) assay. From this experiment, it was confirmed that MAbs produced in this study were PRRSV ORF7-specific and could be used as reliable reagents for type 1/type 2 PRRSV detection.
Keywords
Porcine reproductive and respiratory syndrome virus (PRRSV); Open reading frame 7 (ORF7) gene; Expression; Monoclonal antibody;
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