Objective: To construct a recombinant adenovirus carrying rat prostate-specific membrane antigen (PSMA) gene and to investigate the immune response of mice to it.
Methods: PSMA cDNA fragment was amplified by RT-PCR using total RNA extracted from rat brain tissue. Recombinant plasmid pAd-rPSMA was constructed by using adeasy adenovirus vector system. Packaged recombinant adenovirus Ad-rPSMA were obtained. The expression of rat PSMA gene in HeLa cells was confirmed by Western blot. The recombinant adenovirus were applied to C57 mice and the resulting anti sera against PSMA were detected with ELISA.
Results: Recombinant adenovirus carrying rat PSMA gene was constructed successfully. The virus particle count, A260/A280 value and titer were 1.97 x 10(12) VP/mL, 1.32, 5 x 10(10) IU/mL respectively. Western blot analysis proved the expression of rat PSMA gene in HeLa cells. PSMA-specific sera antibody was produced in mice.
Conclusion: Recombinant adenovirus Ad-rPSMA was constructed and it could induce the mice to produce PSMA-specific antibody.