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A more methodological approach to screen genomes rapidly for immunorelevant antigens

A system was developed to screen large fragments of the PRV genome. First, two transactivator genes IE180 and EP0 were cloned into eukaryotic expression vectors and a set of four fragments, available as cosmid clones and constituting the whole PRV genome, were tested on integrity. The activity of these transactivator products in stimulation the expression of PRV genes was demonstrated. The sequence of PRV cosmids however was not complete and it was not possible to generate the complete PRV genome out of these fragments. Possibly due to the same reason, the expression of genes from the cosmids was not successful.

However, using the substitute large fragments of cloned PRV DNA, the method of screening the genome for immunoprotective antigens was established and optimal ratio of plasmids/transactivator genes for efficient expression were established. Furthermore, baculovirus recombinants expressing IE180 and EPO were constructed and their recombinant antigens for screening the immune response were prepared.

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University of Gdansk
24 Kladki 24
80 822 Gdansk
Poland
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