To construct recombinant adenovirus vector contained human pancreatic duodenal homeobox-1 (PDX-1) so as to study the expression and effect of PDX-1 gene in bone marrow mesenchymal stem cells, human PDX-1 gene was ligated into shuttle vector pAdTrack-CMV. Homologous recombination was performed in BJ5183 bacteria by cotransforming linearized shuttle plasmid with adenovirus backbone plasmid pAdEasy-1.The recombinant plasmid was packaged and amplified in 293 cells.Mesenchymal stem cells(MSCs) were isolated from healthy adult bone marrow.The adenovirus was transfected into MSCs. The recombinant adenovirus vector has been successfully constructed according to the results of sequencing,PCR and enzyme digestion identification.The expression of PDX-1 and insulin in transfected MSCs at 7 days was detected by RT-PCR and immunocytochemistry. The value of insulin secretion was (15.21±3.50 )mIU/L from the transfected cells at 7days.These results showed that PDX-1 gene modified MSCs could be the beta cell replacement for those diabetes patients who need insulin-secreting cells.