Construction and identification of IL-31 recombinant adenoviral plasmid

Objective To construct and identify human IL-31 recombinant adenovirus.Methods IL-31 gene was amplified from recombinant plasmid pGEM-T-IL-31 and cloned into the shuttle plasmid pAdTrack-CMV.The recombinant shuttle plasmid was digested by PmeⅠ,followed by homologous recombination with bone plasmid pAdEasy-1 in BJ5183.Recombinant adenovirus was obtained after being packaged in QBI-293A cells,then identified by RT-PCR and western blot.The titer of virus was analyzed with fluorescent counter.Results In recombinant shuttle plasmid and recombinant adenovirus,a fragment of 500bp detected by RT-PCR and digested by enzyme was identical with that included in Genbank.The titer of recombinant adenovirus reached 2.6×108 pfu/ml.IL-31 mRNA and protein were expressed in QBI-293A cells infected by recombinant adenovirus.Conclusion The recombinant adenovirus containing IL-31 gene was constructed successfully.IL-31 protein could be expressed stably and effectively.This will be helpful for further study of the function of IL-31.