In the previous study in 2013, the authors had cloned Vietnamese chicken interleukin 6 gene. In this study, the authors describe the creation of an recombinant adenovirus containing chIL-6 as an immune stimulation product for poutry. ChIL-6 gene was amplified total RNA extracted from the spleen of Vietnamese chicken cell by RT-PCR. Then ChIL-6 gene was cloned into the shuttle vector pENTR3C that carried attL1 and attL2 sites for site-specific recombination of the entry clone with gateway destination vector to generate a recombinant plasmid pAd/CMV- V5-chIL6. pAdlCMV- V5-chIL6 was exposed by Pad enzyme, the purpose of Pad digestion is to liberate the two ITRs (left and right) of adenoviral genome.