To construct miR-122 overexpression Adenovirus vector,gene including eight miR-122 precursor fragments was inserted into recombinant construct pDC312-cmv. Abilities of the recombinant vector p-8miR-122 to express miR-122 level and anti-HBV activities were analysed by RT-PCR、ELISA、Sourthern blot. Through adenovirus packaging、amplification、purification and titer detection,the pDC312-cmv and p-8miR-122 adenovirus had a high purity,high miR-122 expression levels and significantly anti-HBV activities.