| 摘要 |
PURPOSE: A recombinant adenoviral vector with self-replicating capability of vector DNA without formating of infectious adenovirus particle is provided, which transfers gene-introducing DNA sequence to pharmaceutically active gene; can be used in gene therapy in vivo; and shows improved gene-introducing stability comparing to a vector without self-replicating capability. CONSTITUTION: A process for the preparation of recombinant adenoviral vector comprises steps of: cutting important hetero cDNA; preparing E1 part-eliminated adenoviral mutants; inserting the cut cDNA into revealed plasmid pZS2 adequate to eukaryotic cell; replicating adenoviral vector RP5, and separating; separating virus DNA, cutting, and separating from large 3'-Xbal fraction; performing ligation of the pZS2 with large Xbal fraction, replicating, and performing transfection; dissolving virus-contained cell culture completely, and refining the standard flake; freezing and thawing the flake material repeatedly for 3times, treating at 37°C for 30minutes in a dissolving buffer, and inactivating at 85°C for 10minutes; separating DNA with common phenol/chloroform extracting.
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