PREPARATION OF RECOMBINANT ADENOVIRUS CARRYING A REP GENE OF ADENO-ASSOCIATED VIRUS

摘要

Adenoviral particles are produced by incubating cells containing a helper adenovirus vector and a helper-dependent adenoviral vector including an Adeno-Associated Virus (AAV) rep gene, such as rep78. Cells are provided containing a helper adenovirus vector. A helper-dependent adenoviral vector including an AAV rep gene is introduced into the cells, for instance by infection with infective viral particles. The cells are incubated to produce adenoviral particles containing nucleic acid including the AAV rep gene. Advantageously, cells containing helper adenovirus vector are pre-incubated for 0.5-12 hours, preferably about 4 hours, to allow expression of viral proteins required for adenoviral genome replication before introducing the helper-dependent adenovirus vector including an AAV rep gene.