Recombinant vaccinia virus containing a chimeric gene having foreign DNA flanked by vaccinia regulatory DNA

Abstract

Methods and compositions are provided for the use of vaccinia virus or other poxviruses as vectors for expression of foreign genes. Expression of foreign genes is obtained by combining vaccinia virus transcriptional regulatory sequence with uninterrupted foreign protein coding sequences in vitro to form a chimeric gene. The chimeric gene is flanked by DNA from a non-essential region of the vaccinia virus genome to provide sites for in vivo homologous recombination. These steps are facilitated by the construction of plasmids that contain multiple restriction endonuclease sites, next to the vaccinia transcriptional regulatory sequences, for insertion of any foreign protein coding sequence. Transfection procedures are used to introduce the DNA into cells where homologous recombination results in the insertion of the chimeric gene into a non-essential region of the vaccinia virus genome. Infectious vaccinia virus recombinants are distinguished or selected by expression of the foreign gene, loss of activity of a vaccinia virus gene, or by DNA—DNA hybridization. Expression of the foreign gene is obtained by infecting cells or animals with the recombinant vaccinia virus. Examples are provided to show expression of prokaryotic, RNA virus and other DNA virus genes in vaccinia recombinants.

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