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| Expression of truncated NSP2 protein of porcine reproduction and respiratory syndrome virus in E.coli and the development of monoclonal antibodies against NSP2 protein |
| 王海燕 WANG Hai-Yan JIANG Ping JIANG DU Yi-jun DU LI Yu-feng LI LI Jun-xing LI SHENG Fang SHENG |
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Abstract The NSP2 gene of porcine reproductive and respiratory syndrome virus （PRRSV）S1 strain was partly amplified and cloned into a prokaryotic expression vector pGEX-6P-1，and a fusion protein GST-tNSP2 with molecular weight of 50 kD was expressed in E. coli. The purified GST-tNSP2 protein showed a strong reaction with the PRRSV-positive sera in Western-blotting assay. BALB/c mice were immunized with the purified protein, and the splenocytes of the immunized mice were fused with murine myeloma cells SP2/0. After subcloning by 3 times, two hybridoma clones which produced McAbs steadily were screened by ELISA, named 3H3 and 2B5. They all reacted strongly with the PRRSV S1 infected Marc-145 cells in IFA, but not with the PRRSV SY0608 strain. Both of the McAbs were belong to IgG1 isotype, and their light chain were Κ. It indicated that the expressed GST-tNSP2 protein and McAbs could be used for identification of PRRSV isolates and functional analysis of NSP2.
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Received: 08 November 2007
Published: 25 February 2008
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Corresponding Authors:
JIANG Ping JIANG
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