Min  Li, Yuanping  Wang, Nianli  Zou, Sanjie  Cao, Xintian  Wen, Yong  Huang, 
    The Polyclonal Antibody Against Chicken Interleukin-6 Prepared by Immunization of Recombinant Eukaryotic Plasmid Can React Efficiently with its Prokaryotic Expression Protein,
    Journal of Animal and Veterinary Advances,
    Volume 9,Issue 21,
    2010,
    Pages 2679-2684,
    ISSN 1680-5593,
    javaa.2010.2679.2684,
    (https://makhillpublications.co/view-article.php?doi=javaa.2010.2679.2684)
    Abstract: Interleukin-6 (IL-6) is a multifunctional cytokine which has a wide range of activity in animals and has an impact on almost all immune system cells. To produce specific antibody against chicken IL-6 and detect its activity, its complete Open Reading Frame (ORF) sequence was amplified and inserted into the eukaryotic expression vector pcDNA3.1 (+) to get recombinant eukaryotic plasmid pcDNA-ChIL-6 and BALB/c mouse were immunized with this plasmid to produce polyclonal antibody against Chicken IL-6. Then the mature protein coding genes of chicken IL-6 was cloned and inserted into the prokaryotic expression vector pET-32a (+) to get recombinant prokaryotic plasmid pET-ChIL-6, the expression of this fusion protein was successfully induced by Isopropy 1-β-D-thiogalactoside (IPTG) and the recombinant protein was further purified by NTA His•Bind column. The purified recombinant protein could react positively with polyclonal antibody against pcDNA-ChIL-6 in Western-blot and indirect ELISA.
    Keywords: Chicken IL-6;eukaryotic expression;prokaryotic expression;polyclonal antibody;Western-blot;indirect ELISA