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Updated: Jul 18, 2026
An Efficient Method for Adenovirus Production
Published on: June 10, 2021
[Construction and expression of recombinant adenovirus expression vector of beta-defensin-2]
Zhuo Shi1, Qiang Shu, Zheng-yan Zhao
1The Affiliated Children's Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China.
Objective:
To explore the possibility of stable expression of cationic peptide beta-defensin-2 in eukaryocytes with adenovirus vector.
Methods:
The rat beta-defensin-2 (rBD2) gene was cloned at the downstream of CMV promoter of the adenoviral shuttle plasmid pShuttle-CMV. Then pShuttle-CMV-rBD2 was transformed into E. coli BJ5183-AD-1, in which recombination occurred between plasmids and pAdEasy-1 to construct pAdEasy-rBD2. After confirmation by endonuclease, linear pAdEasy-rBD2 was transformed into 292 cells to obtain packaged adenoviral expression vector, which was used to infect cos-7 cells and to establish respiratory adenovirus infection model of rat. The in vivo and in vitro expression activity of recombinant adenovirus was detected by Western blot and immunohistochemistry.
Result:
The inserted DNA of pShuttle-CMV-rBD2 consisted of rat beta-defensin-2 gene. The pathological effect of infected cells, electronic microscopic observation and PCR showed that the recombinant adenovirus vector was constructed successfully. The concentration of the adenovirus was 10(9) PFU/ml. The vector expressed rat beta-defensin-2 efficiently in vivo and in vitro.
Conclusion:
The recombinant adenovirus vector can express cationic peptide beta-defensin-2 in eukaryocytes.
