Construction and identification of recombinant adenovirus carrying human hypoxia-inducible factor-1α gene

Objective To construct the adenovirus vector carrying the recombinant human hypoxia-inducible factor-1 and amplify the adenovirus vector in 293 cells. Methods The recombinant human HIF-1α gene was obtained from constructed plasmid pc-DNA4-rhHIF-1α by RT-PCR and then inserted into linearized pEGFP-C1 plasmid.The EGFP-rhHIF-1α gene segment was amplified by RT-PCR from pEGFP-rhHIF-1α vector and cloned into the plasmid PUC18.After having been screened,the constructed pUC18-EGFP-rhHIF-1α plasmid was digested with restriction endonucleases and cloned into the shuttle plasmid PDC316 to form PDC316-rhHIF-1α vector.The PDC316-rhHIF-1α plasmid was cotransfected with genomic plasmid pBHGloxΔ1,3Cre into 293 cells to package the recombinant adenovirus.The recombinant adenovirus was transfected into SG-7901 cells,and the green fluorescence protein expression was detected. Results Recombinant adenoviral vector Ad-rhHIF-1α was constructed successfully,and confirmed by restriction enzyme digestion and GFP expression. Conclusion The recombinant adenoviral vector carrying rhHIF-1α can be successfully constructed.