China Animal Husbandry & Veterinary Medicine ›› 2009, Vol. 36 ›› Issue (10): 39-43.

• 生物技术 • Previous Articles     Next Articles

Expression and Development of Monoclonal Antibody against Nsp2 Gene Fragments of Highly Virulent PRRSV JXA1 Strain

HU Hong-hui1,2, CAO Zhen2, ZHAO Tie-zhu2, WANG Bin3, QIN Ya-man2,
YANG Lu2, WANG Chuan-bin2, CHEN Xi-zhao2, TIAN Ke-gong2   

  1. (1.College of Veterinary Medicine, China Agricultural University, Beijing 100193, China; 2.China Animal Disease Control Center, Beijing 100125, China; 3.Beijing Animal Health Inspection Institute, Beijing 100044, China）
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-10-20 Published:2009-10-20
  • Contact: TIAN Ke-gong

Abstract: Analyzing PRRSV gene Nsp2 by biological software, four gene fragments of Nsp2 were successfully amplified by RT-PCR and cloned from the PRRSV JXA1 strain. Their expressing plasmids were constructed by inserting the target gene fragments into pGEX-6P-1 vector and were induced with IPTG. Subsequently, the collecting and purifying proteins were identified. Using these proteins, we established an indirect ELISA for bolting McAbs. We immunized the BALB/c mice with PRRSV vaccine. After the final immunization with virus, spleen cells of mice fused with myeloma cells SP2/0 were filtered by culturing selectively, diluting finitely. As a result, an anti-Nsp2 protein McAb was obtained and named as 6B8. It is only with reaction to Nsp2F1. The ELISA titers of 6B8 supernatants and ascites were 210 and 106 respectively. In this study, firstly we successfully cloned and expressed Nsp2 gene fragments of PRRSV JXA1, secondly an anti-Nsp2 protein McAb was obtained. It is a basic research for analyzing the function of Nsp2 and establishing related diagnosis method.

Key words: PRRSV JXA1; Nsp2; McAb

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