Construction of recombinant adenovirus and its transgenic expression in cultures of Corti's organ and spiral ganglion cells

Objective To construct recombinant adenovirus carrying the gene for green fluorescent protein(Ad-GFP) and observe the expression of GFP in the cultures of Corti's organ and spiral ganglion cells of the neonatal rat.Methods The Ad-GFP was constructed by homologous recombination in bacteria.The expression,distribution of GFP,and the effect of adenovirus on the cultures of Corti's organ and spiral ganglion cells of the neonatal rat were observed at different time points after transfection.Results The constructed recombination was proved to be correct by the agarose gel electrophoresis after restriction digestion.The cultures of Corti's organ and spiral ganglion cells of the neonatal rat began to express GFP gene 12 h after transfection with virus.Gene expression reached a peak level after 24 h and persisted for 1 week.The structure and growth of Corti's organ and spiral ganglion cells were not significantly changed after transfection.Conclusion The recombinant Ad-GFP has been constructed successfully.The adenovirus can mediate GFP expression in the cultures of Corti's organ and spiral ganglion cells of the neonatal rat without great influence on the cultures.Therefore,the constructed recombinant adenovirus may have important value for gene therapy of the inner ear.