[Show abstract][Hide abstract] 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-ChlL-6 in Western-blot and indirect ELISA.
Preview · Article · Dec 2010 · Journal of Animal and Veterinary Advances