Construction Of Human Adeno-associated Virus Type2 (AAV-2) Based Vectors, Expression Of Foreign Genes In The Vectors And A Study On The Recombinant AAV Packing System

In this experiment, based on vectors pSSV9 and pSSV9-int, which contain the genome of wild type human adeno-associated virus type 2 (AAV-2), We constructed several AAV vectors. The vectors are pSSV9/P40-LacZ(+), pSSV9/P40-LacZ(-), pSSV9/ MuLV-neosv and pSSV9/MuLV-neosv -IL2. We also constructed a nonviral vector pGEM-3zf/P40-LacZ(1).After introducing the vectors into 293 cells and A549 cells by lipofectin-mediated transfection, the biological activity of the gene expression products inclucling β -galactosidase,neomycin resistant substance and human interleukin 2, were identified.In A549 cells and 293 cells, transcient expressions of LacZ gene in the recombinant AAV vectors pSSV9/P40-LacZ(+), pSSV9/ P40-LacZ(-), and in the non-viral vector pGEM-3zf/ P40 -LacZ(1) were done. The results show:① In these mammalian cells, gene expression in the AAVvectors changed chronologically. The expression occured soon(12-24 hour) after the transfection, boomed to its maximum (in 24 hours), thereafter, dropped quickly (in 24-72 hours), and lasted a longer time at low levers.② A change in the structure of AAV itr influenced the...