Objective  To construct the human PDX1 and NKX6.1 doublegene adenovirus expression vector using the pADxsi system.   Methods  pShuttle-GFP-CMV-NKX6.1 was constructed by cloning the target gene NKX6.1 into pShuttle-GFP-CMV. pShuttle-CMV-PDX1/CMV-NKX6.1 was constructed by cloning PDX1 into pShuttle-CMV-GFP/CMV-NKX6.1 to replace GFP. The pADxsi-CMV-PDX1/CMV-NKX6.1 was constructed by transferring CMV-PDX1/CMV-NKX6.1 to ADxsi backbone plasmid vector from pShuttle-CMV-PDX1/CMV-NKX6.1. Then

the pADxsi-CMV-PDX1/CMV-NKX6.1 adenovirus vector was amplified in 293 cells and viral titer was measured. Human placental liver mesenchymal stem cells were infected with adenoviruses.   Results  Restriction enzyme analysis and PCR showed that the PDX1 and NKX6.1 double-gene adeno-virus expression vector was successfully constructed

which could infect the human placental liver mesenchymal stem cells and was specifically located in nuclei.   Conclusion  Human PDX1 and NKX6.1 double-gene adenovirus expression vectors can be successfully constructed using recombinant techniques of the pADxsi system.