Objective:To construct the recombinant adenovirus expression vector containing the transcription factor ZNF191 and to evaluate its potential role for gene therapy of tumor.Methods:The ZNF191 cDNA was amplified by RT-PCR using template mRNA isolated from the HEK293 cells.The cDNA fragments were cloned into the shuttle vector pAdTrack-CMV.The product pAdTrack-ZNF191 was linearized by PmeⅠ for homologous recombination with pAdEasy-1 in pAdEasy-1 competence bacteria.The positive clone was identified by enzyme digestion

PCR analysis and DNA sequence analysis.After linearization by PacⅠ

the recombinant adenovirus DNA plasmid pAdEasy-ZNF191 was transfected into 293 T cells for packaging and amplifying Ad-ZNF191

which was further identified by PCR analysis and DNA sequence analysis.Western blotting was used to detect the expression of ZNF191 protein in the MCF-7 cells infected with the adenovirus.Results:The pAdTrack-ZNF191 and pAd-Easy-ZNF191 plasmids had been successfully constructed and they were verified by PCR analysis

enzyme digestion and DNA sequence analysis.PCR analysis and DNA sequence analysis confirmed successful packaging of the recombinant adenovirus Ad-ZNF191 in 293T cells.ZNF191 protein expression increased significantly in the MCF-7 cells after infection by the recombinant virus.Conclusion:We have successfully constructed the recombinant adenovirus Ad-ZNF191

which provides a basis for investigating the biological function of ZNF191 and its potential for gene therapy.