Replication stable and RNase resistant chimeras of pestivirus with insertion in 3′ nontranslated region (3′NTR)

A vector comprising:

• DNA sequence of a chimeric viral RNA sequence comprising:
• vector DNA sequence, a first DNA sequence complementary to a first Pestivirus RNA sequence, and a second DNA sequence complementary to a second RNA sequence, wherein the second RNA sequence is inserted within a variable region of a 3′ non translated region between a SLSTOP stem-loop structure and a SLII stem-loop structure in the 3′ non translated region, NTR, of the first Pestivirus RNA sequence, and wherein the sequence of a chimeric RNA virus generated by transcription of the DNA vector is stable in replication, and forms a RNase resistant viral particle, and wherein the first virus RNA sequence is from a bovine viral diarrhea virus (BVDV).

A culture of cells containing RNA virus comprising:

• host cell harboring chimeric first and second RNA virus sequence, wherein the first RNA virus is Pestivirus and a second RNA sequence is inserted in a variable region of a 3′ non translated region between a SLSTOP stem-loop structure and a SLII stem-loop structure in the 3′ non translated region, NTR, of the first Pestivirus RNA sequence, wherein the chimeric RNA viral sequence is stable in RNA replication, and ribonuclease resistant viral particles comprising the first and second RNA virus sequences when cultured, and wherein the first virus RNA sequence is from a bovine viral diarrhea virus (BVDV).