To construct a recombinant adenovirus vector carrying the human GFAP promoter and p27 gene and identify its effects on cell growth and proliferation after transfecting into astrocytes. Methods: The original pGFAP-IRES2-EGFP-p27 plasmid in the laboratory was transformed, amplified, extracted, digested, identified and sequenced by agarose gel electrophoresis. By designing primers and conducting PCR amplification, GFAP promoter and p27 gene fragments were obtained, and then exchanged into linearized expression vectors GV269 and pDC315-GFAP-EGFP, respectively. Finally, pDC315-GFAP-p27-EGFP was constructed. Transfect it into HEK293 cells and package it into recombinant virus particles. After repeated amplification of several generations in HEK293 cells, purify it and detect its titer. Observe cell growth and fluorescence expression after transfecting astrocytes. Detect the cell cycle and apoptosis ratio of astrocytes after transfection, as well as the expression of p27. Result: After enzyme digestion identification and sequencing, the eukaryotic expression vector pGFAP-IRES2-EGFP-p27 was successfully identified. After PCR identification and sequencing, the recombinant adenovirus overexpression vector pDC315-GFAP-p27-EGFP was successfully constructed. After packaging, Ad-GFAP-p27-EGFP showed EGFP expression and cytopathic effects, and was able to specifically express green fluorescent protein in astrocytes. According to flow cytometry analysis, 72 hours after transfecting astrocytes, compared with the control group, the proportion of cells in the S phase decreased in the transfection group, while the proportion of cells in the G1 and G2 phases increased and the proportion of apoptosis increased (P<0.05). On the 5th day after transfection, the expression level of p27 protein in astrocytes significantly increased compared to the control group (P<0.05). Conclusion: A recombinant adenovirus overexpression vector carrying the human GFAP promoter and p27 gene was successfully constructed using the adenovirus packaging system AdMax. It can specifically transfect astrocytes to overexpress p27, and has a significant inhibitory effect on the growth and proliferation of astrocytes.