Cloning, Expression And Detection Of Biological Activity Of The Chicken IL-18 Gene

Domestic infective diseases which cause the tremendous economic losses to poultry industry, are the most large threat of poultry industry,in the useness of the traditional immunity there still exist some questions, for instance, the potential safe problem of weak poisonous vaccine; Deactivation vaccine although have effective security, it renders a restriction to immunity, especially cell immune restricted function. Genetic engineering inferior unit vaccine and DNA vaccine have potentiality of using, but it is also necessary to improve their immunity, especially cell's immune effect.IL-18 is a kind of the powerful inducer of IFN-γ, which has the important immune regulating function, it has been one of the focuses in the cytokine's studying all the time since it was found, and it may become one of the important adjuvant of cell immunity, in order to improve chicken body's anti-infective activity, or strengthen the immune effect of the deactivation vaccine or genetic engineering vaccine.This research, according to the gene sequence of ChIL-18 which GenBank issued, designed one pair primer on both sides of its coding region, the chicken of Gushi k, Gushi chicken and Hailan, etc. several variety chicken on the about 60th age which were breeded by a large scale in Henan, abstracted the total RNA completely as the template from its spleen lymphocyte of activated with mitogen PHA, RT-PCR all amplificated genetic fragments about 600bp, and make it cloned to pGEM-T Easy Vector, constructured the recombinant plasmid pGEM-ChIL-18, then it identificated by PCR, enzyme digestion and DNA sequencing to the whole gene of ChIL-18, which indicate by contrast: the three whole gene sequence and their deduced amino acid of ChIL-18 array 597bp, which include one open-reading frame, and encode 198 amino acid, and array 29 amino acid which is signal peptide in N end, ChIL-18 gene sequence accords with that issued by Schneider, etc. (2000) fundamentally; IL-18 gene have variety in species, blood reason relation homology is high, establish the foundation as immunity adjuvant to apply in research of immunization vaccination.On this basis, mChIL-18 gene which derived from pGEM-ChIL-18 was digested...