Baculovirus expression system

Method for producing a recombinant baculovirus of which the genome comprises one or more transgene(s) each encoding a protein maturation enzyme and n transgenes each encoding a polypeptide of interest, said method comprising the steps of:

• a) Preparing, in an insect cell, a recombinant baculovirus genome capable of replicating which comprises one or more transgene(s), each encoding a protein maturation enzyme and n transgenes each encoding a polypeptide of interest, by homologous recombination between:
• a1) a replication deficient baculovirus genome in which n genes essential for viral replication are non-functional and which comprises one or more transgene(s), each encoding a protein maturation enzyme, and
• a2) n transfer vectors each comprising:
• i) a nucleotide sequence enabling to restore the function of one of the n non-functional genes essential for viral replication,
• ii) one of the n transgenes encoding a polypeptide of interest,
• the set of nucleotide sequences i) of the n transfer vectors being capable of restoring the replication of the replication deficient baculovirus genome,
• n being an integer at least equal to 2;
• and
• b) Generating a recombinant baculovirus in an insect cell which comprises the recombinant baculovirus genome obtained at step a).

Set of homologous recombination elements comprising:

• a) a replication deficient baculovirus genome in which n genes essential for viral replication are non-functional and which comprises one or more transgene(s) each encoding a protein maturation enzyme;
• b) n transfer vectors each comprising:
• i) a nucleic acid sequence enabling to restore the function of one of the n non-functional genes essential for viral replication,
• ii) a transgene encoding a polypeptide of interest,
• n being an integer at least equal to 2.