Pharmaceutical composition comprising the viral gene transfer and attenuated virus vaccination, and production methods of the virus

摘要

(57) RNA polymerase I transcription in vivo in cells that have been DNA transfected [summary] temporary, expression of influenza vRNA molecules encoding chloramphenicol acetyl transferase (CAT) in the antisense direction it was used for. Co-infection of influenza virus, served for transcription conversion of negative strand vRNA to the positive-strand viral mRNA molecules expressing the CAT activity, to provide protein and other viral RNA polymerase. Nucleotide exchange, and the system was used for analysis by deletions and insertions of both terminal segments of the vRNA sequence which cooperatively constitute the vRNA promoter structure. Variants of several with the expression rate that has been greatly enhanced in the wild-type level or higher has been constructed, but it also, is packaged in progeny virus, can be serially passaged. Data obtained for the mutation in the promoter of various factors, to support the model of the double-stranded vRNA promoter structure that is consistent in the initiation of RNA synthesis and the binding of the viral polymerase. Preparation of attenuated viruses for vaccination purposes, including recombinant segment with a single promoter promoting mutation for overexpression of the gene product of foreign or myself, of its overexpression at the same time In order to work on reducing the helper virus RNP number of segments. Moreover, the same virus was passaged through stages of a ribozyme cleavage acting on one segment of a helper virus, which would lack the structure and function of this virus at a high rate from the progeny virus. Attenuated viruses obtained will not be able to interact with its target cells only in one round of abortive infection, to produce progeny virus.