Abstract: Abstract Objective：To construct a lentiviral vector which contains short hairpin RNA(shRNA) interfering sequence aiming at the target of human Papillomavirus 18E6 (HPV18E6) oncogene.Method：Based on the information of HPV18E6 oncogene,design and synthesize shRNA interfering sequence,recombinate into pGCSIL-GFP plasmid to construct a recombination plasmid pGC-shRNA which contains shRNA interfering sequence and apprase it；Recombination plasmid and packaging plasmid pHelper 1.0 and pHelper 2.0 co-transduce the 293T cell to package the lentiviral vector,collects the lentiviral vector solution,concentrates and purifies it；The lentiviral vector solution after concentrating and purifying transduce 293T cell to determine the concentration of the lentiviral vector solution.Results：Indexing shRNA of interfering sequence aiming at the target of HPV18E6 oncogene with human genes in BLASTER,there is no homogeneous,the second constructure of the transcripts of interfering sequence can conform short hairpin；The recombination plasmid pGC-shRNA was obtained with the correct sequence in the insert after the appraisal；After packaging and collecting and concentrating the lentiviral vector successfully, the concentration of lentiviral vector solution is 3x108TU/ml.Conclusion：The lentiviral vectors of HPV18E6-RNAi-LV was constructed successfully,which provides practical tool for later study.

Key words: HPV18E6, short hairpin RNA, recombination plasmid, lentiviral vector, RNA interfere