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Updated: May 7, 2026
An Efficient Method for Adenovirus Production
Published on: June 10, 2021
[Construction and identification of a recombinant replication-defective adenovirus vector encoding p53AIP1 and its
Xiaoliang Lin1, Yunbo Jiang, Qian Zhao
1Department of Urology, General Hospital of PLA, Beijing 100853, China.
Objective:
To construct a recombinant replication-defective adenovirus vector carrying p53AIP1 (p53-regulated apoptosis-inducing protein 1) gene and observe its expression in human HeLa cells.
Methods:
Specific primers were designed according to p53AIP1 gene sequence and inserted specific enzyme cutting sites. P53AIP1 gene was amplified by PCR and cloned into the adenovirus shuttle plasmid pDC316 to construct the recombinant vector pDC316-p53AIP1, which was co-transfected with helper plasmid pBHGloxδE1, 3Cre into HEK293 cells by Lipofectamine(TM); 2000. The recombinant replication-defective adenovirus (Ad-p53AIP1) was generated by means of homologous recombination of the two plasmids in HEK293 cells with the Cre-loxP recombinase system and harvested after 12 days. Ad-p53AIP1 and Ad-null were respectively transfected into HeLa cells at MOI=100. Then the expression of p53AIP1 gene was detected by Western blotting.
Results:
pDC316-p53AIP1 was constructed successfully. The new constructed vector was confirmed by PCR analysis, double enzyme digestion and DNA sequencing. The results were in conformity with the expected. Western blotting demonstrated that the target p53AIP1 proteins were effectively expressed in the transfected HeLa cells.
Conclusion:
The recombinant adenovirus vector of Ad-p53AIP1 was successfully established, and it was proved to have a strong ability of infectivity.
