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KMID : 1023520090320010019
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Korean Journal of Veterinary Service
2009 Volume.32 No. 1 p.19 ~ p.25
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Expression of ORF6 gene of porcine reproductive and respiratory syndrome (PRRS) virus
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Bae Su-jung
Kim Jin-Won
Yoon Young-Sim
Kang Shien-Young
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Abstract
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Porcine reproductive and respiratory syndrome (PRRS) virus is the etiological agent of diseases characterized by reproductive losses in sows and respiratory disorders in piglets. The PRRS virus is a small enveloped virus containing a positive-sense, single-stranded RNA genome. In the present study, ORF6 gene of Korean PRRS virus isolate, CNV, was cloned and expressed in baculovirus expression system. The ORF6 gene and expressed protein in the recombinant virus were confirmed by PCR/indirect fluorescence antibody (IFA) test and Western blotting, respectively. The recombinant protein with a molecular weight of approximately 24KDa was confirmed by Western blotting using His6 and PRRS virus-specific antiserum. Expressed ORF6 protein was applied for IFA to detect antibody against PRRS virus using field porcine sera. However, the sensitivity and specificity of developed IFA using expressed ORF6 protein were considerably low compared to those of commercial ELISA kit. This results suggest that IFA using expressed ORF6 protein could not be used as a diagnostic test for PRRS virus infection without further improvements.
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KEYWORD
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PRRS virus, ORF6, Baculovirus expression
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