Establishment of human papillomavirus type 16 E6/7 transgenic mouse model

Objective To construct pCEP4-human papillomavirus type 16(HPV16)E6/7 gene eukaryotic expression plasmid,and to set up HPV16 E6/7 transgenic mouse model by microinjection. Methods Genetic recombination technique was used to construct eukaryotic expression plasmid pCEP4-HPV16 E6/7.The recombinant plasmid was microinjected into male pronuclei of donor mice,which were thentransplanted into fertilized ovum of pseudopregnant mice.Totally,129 founder mice were born. Of them,5 were confirmed by PCR to be transgenic mice.As Southern blot showed,4 of the 5 transgenic mice yielded specific fragments similar to the positive plasmid,which suggested that E6/7 gene was integrated into the genome of these 4 mice.The founder mice were mated with normal FVB mice,and pCEP4-E6/7 gene was detected by PCR in their descendants.The results confirmed that the exogenous gene had been stably inherited by their descendants.Epithelial dysplasia of skin tissue was observed in 2 of 4 5-months-old trans- genic mice.Results The eukaryotic expression plasmid pCEP4-HPV16 E6/7 was constructed successfully, and epithelial dysplasia of skin tissue was observed in its transgenic mice.Conclusion HPV16 E6/7 trans- genic mouse model,with the capability of propagating and passaging,is constructed successfully.