›› 2014, Vol. 41 ›› Issue (9): 20-24.

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Prokaryotic Expression of Duck Retinoic Acid inducible-gene Ⅰ and Preparation and Identification of Monoclonal Antibody against the Protein

ZHOU Chang-liang1, ZHANG Ya-chun1, WANG Wei1, LI Yue1, ZHAO Ying-hui1, ZANG Feng-xia1, RAN Duo-liang2, MENG Qing-wen1, CHEN Hong-yan1   

  1. 1. National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China;
    2. College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830000, China
  • Received:2014-04-10 Online:2014-09-20 Published:2014-09-24

Abstract: In order to prepare the monoclonal antibodies against duck retinoic acid inducible-gene Ⅰ(RIG-Ⅰ) full-length protein, the N-terminal a (1 to 900 bp)and b (751 to 1650 bp) segments of RIG-Ⅰ gene were amplified from cDNA of duck spleen and then cloned into the prokaryotic expression vector pET-30a. The recombinant plasmid was transformed into BL21 cells and expressed by IPTG inducing,then immuned BALB/c mice by the protein, and fused the spleen lymphocytes with SP2/0 myeloma cell, detected by indirect ELISA. 17 strains hybridoma cell lines which had good reactivity with duck RIG-Ⅰ protein were screened; the titer of monoclonal antibody supernatant was 1:512; 2 strains monoclonal antibody 10H7C3 and 2A10A2 had good reactivity with duck RIG-Ⅰfull-length protein detected by indirect immunofluorescence (IFA) and Western blotting. The specific murine anti-duck RIG-Ⅰ monoclonal antibodies that prepared in the study laid a foundation for further research of RIG-Ⅰ gene function.

Key words: RIG-Ⅰ; monoclonal antibody; prokaryotic expression

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