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To construct and identify further a recombinant of Adeno-associated virus and interferon-gamma for gene therapy, the full-length IFN-%γ%cDNA containing signal peptide was amplified by PCR, and then cloned into the pUC18. After screening, the fragment from the positive clone was then subcloned into pwp19. After the correct recombinant was identified by digestion with SacI and BamHI, it was transfected into lymphocyte cell line H9 mediated by calcium phosphate, and the expression of IFN-%γ% was detected by RT-PCR and ELISA. The result showed that the IFN-%γ% were expressed in the H9 cells transfected with pwp/IFN-%γ%. The so constructed recombinant plasmid pwp19/IFN-%γ% containing the full-length IFN-%γ% gene was expressed in mammalian cells.
To construct and identify further recombinant Adeno-associated virus and interferon-gamma for gene therapy, the full-length IFN - % γ % cDNA containing signal peptide was amplified by PCR, and then cloned into pUC18. screening, the fragment from the positive clone was then subcloned into pwp19. After the correct recombinant was identified by digestion with SacI and BamHI, it was transfected into lymphocyte cell line H9 mediated by calcium phosphate, and the expression of IFN - % gamma % was detected by RT-PCR and ELISA. The result showed that the IFN - % γ % were expressed in the H9 cells transfected with pwp / IFN - % γ %. The so constructed recombinant plasmid pwp19 / IFN - % γ % containing the full-length IFN - % γ % gene was expressed in mammalian cells.