Construction of recombinant adenovirus of mouse A20 gene and identification of cochlear hair cells

Objective To construct the recombinant adenovirus of mouse A20 gene and to observe its ability to infect hair cells of guinea pig inner ear.Methods The A20 gene fragment was cloned into the shuttle plasmid pAdTrack-CMV to form the transfer vector by the method of homogenous recombination in bacteria.Then the recombinant adenovirus was transfected into 293 cells using liposome DOTAP.The target gene was detected by polymerase chain reaction(PCR).The titer and infection rate of the recombinant adenovirus were determined using the green fluorescent protein(GFP)expression in the shuttle plasmid.Results Restriction endonuclease and PCR analyses confirmed that the A20 gene was successfully inserted into the adenovirus vector.The titer of the recombinant adenovirus was 5×109 pfu/mL.The adenovirus had a strong effect on hair cells of inner ear.Conclusion The recombinant adenovirus containing A20 gene is successfully constructed by the method of homogenous recombination in bacteria.