Method for the production of recombinant virus, dna constructs, recombinant virus and vaccine compositions

Abstract

The purpose of the present invention is the production of recombinant virus through the cloning and expression of sequences of nucleotides encoding whole or part of heterologous proteins, through the following method: (a) modification of the heterologous nucleotides sequences in such way that when cloned and expressed in a vector virus, the 5' region nucleotides present in the 5′ edge of the gene NS1 of this vector virus or of other virus or equivalent functional sequences, and in its 3′ region, the correspondent genome region in the whole or part of the domains of the stem and anchor of the protein E of the vector virus or equivalent functional sequences, and not comprising the structure and the replication of the mentioned vector virus; (b) insertion of the modified heterologous sequences in (a) in the intergenic region at the structural protein E level and of the nonstructural NS1 vector virus; (c) obtaining the non pathogenic recombinant virus with immunologic properties, having the heterologous sequences integrated in the viral genome according to the insertion described in (b) and, then expressing the heterologous antigen in such way that it can induce an appropriate immune response. The present invention is also addressed to vaccine compositions to immunize against the Flavivirus and/or other pathogens.