Development of new expression vector based on Pseudorabies virus amplicons: application to human insulin expression

Pseudorabies virus (PrV), a herpesvirus from the Alphaherpesvirinae subfamily, is suitable for amplicon vector replication and packaging into virions, with helper virus for trans replication and cleavage-packaging functions. PrV amplicon vectors were developed in a bacterial plasmid construction using PrV ori(s) and pac signals as the required cis elements. Human insulin cDNA was then cloned in the amplicon vector for human proinsulin expression. In the same construction, green fluorescent protein was used as a marker. PrV amplicons may have several advantages over herpes simplex virus type 1 (HSV1) amplicons in human gene therapy because it can infect human cells in vitro and in vivo, it is not pathogenic for primates and there is no pre-existing immunity and risk of recombination with latent PrV as occurs with HSV1.